infocus Magazine - Issue 73, March 2024

ISSUE 73 MARCH 2024

The Scanning Microscope and Leeuwenhoek’s Legacy RMS Summer Studentship Reports 2023 The RMS Mentoring and Application Scheme: Nurturing Microscopists across the world Reflections on the RMS Diploma: Study, Mentorship, and Oversight

Plus...News, Calendar, Reviews, Reports 1

ISSUE 73 MARCH 2024

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contents features 4

The Scanning Microscope and Leeuwenhoek’s Legacy Professor Brian Ford, Hon FRMS

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RMS Summer Studentship reports 2023 Lakshana Baheerathan, Mohamed Ghali, Miriam Czech, Hio U Lao, Rebecca Harry

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The RMS Mentoring and Application Scheme: Nurturing Microscopists across the world Joëlle Goulding and Georgina Fletcher

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Reflections on the RMS Diploma: Study, Mentorship, and Oversight Tim Young, Joëlle Goulding, Susan Brooks

regulars 10 38 45 48 61 65

Calendar Journal of Microscopy New Member Welcome Office News Company News New Products

reports and other features 14 36 42 78 81 82

Global BioImaging workshop - Cuernavaca, México Register now for elmi2024!

NMDC 2023, Paestum, Italy RMS Diploma: Meet our latest Candidates!

MRS Fall Meeting 2023, Boston, Massachusetts, USA From the archives: A ‘double-take’ from 1989

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MAGA Z I N E

infocus is the Magazine of the Royal Microscopical Society (RMS) – the only truly international microscopical society. The RMS is dedicated to advancing science, developing careers and supporting wider understanding of science and microscopy. infocus Magazine 37/38 St Clements Oxford, OX4 1AJ, UK Tel. +44 (0)1865 254760 Email: infocus@rms.org.uk Website: www.infocus.org.uk Scientific Editor Leandro Lemgruber, University of Glasgow, UK Editor Owen Morton Tel. + (0)1865 254763, Email: editor@infocus.org.uk Editorial Board Susan Cox, King’s College, London, UK Rebecca Higginson, Loughborough University, UK Laura Fumagalli, University of Manchester, UK Myfanwy Adams, John Innes Centre, Norwich, UK Maadhav Kothari, Zeiss Microscopy, UK Hilary Sandig, Cancer Research, UK Trevor Almeida, University of Glasgow, UK Advertising Email: advertising@infocus.org.uk ISSN: 1750-4740 © 2024 Royal Microscopical Society infocus is published four times per year by the RMS. Designed and produced

by Creative Design. Reproduction in whole or in part without permission from the RMS is forbidden. Views expressed in the Magazine are those of the individual contributors and do not necessarily reflect those of the RMS.

RoyalMicroscopicalSociety @RoyalMicroSoc @RoyalMicroSoc

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ISSUE 73 MARCH 2024

FROM THE SCIENTIFIC EDITOR Dear Readers, Happy New Year! I hope the first months of 2024 have treated you well – whether you’ve been experiencing the thick of winter in the northern hemisphere or something a little warmer in the south. It is a pleasure to bring to you the first issue of 2024.

This issue marks our first anniversary of infocus being a fully online publication. It has been a year of learning and new adventures for us all in the infocus team. We hope to continue bringing you the best possible range of content throughout 2024. Speaking of learning curves, we have a great piece on the RMS Diploma - a flexible, portfolio-based qualification, designed to be similar to a Masters degree.We get to hear three different perspectives on the programme – from the Diploma candidate, the mentor and from the Chair of the RMS Qualifications committee.

Another important strand of the RMS’s educational activities is the annual Summer Studentship scheme for undergraduates in their second year of university study.The students carry out projects involving microscopy or image analysis in physical or life sciences, submitting a report at the end. In this issue, we are delighted to publish all five reports from the ‘class of 2023’. Mohamed Ghali (University of Nottingham) details his project on the mechanical meniscus correction in phase contrast microscopy; Hio U Lao (Strathclyde University) describes the use of Raman scattering microscopy to study the uptake and distribution of Bruton’s tyrosine kinase inhibitors; Rebecca Harry (University of Oxford) details her project using microtubule depolymerisation assay to study BRCA2 and MCAK interactions; Miriam Czech (King’s College London) reports on her project evaluating different metrics used in deep learning for microscopy; and Lakshana Baheerathan (Durham University) describes her use of high-resolution scanning electron microscopy to image the LINC complex at the nuclear envelope. It is always fantastic to see young people getting involved in microscopy.

A couple of years ago, the RMS launched its Mentoring Working Group with the aim of helping scientists develop new skills and receive specialist support towards career development. In a further education-themed article, some of our current mentees and mentors provide their feedback and describe their experiences.

Finally, anyone with an interest in the history of microscopy is sure to be captivated by the latest article from RMS Honorary Fellow Brian Ford, who brings us an amazing story on the use of scanning electron microscopy to study and characterise Leeuwenhoek’s microscopes, 300 years after his death.

I hope you enjoy everything this issue has to offer, and look forward to seeing as many of you as possible at elmi2024 in June – or one of our other RMS events later this year!

Slàinte!

Leandro Lemgruber

COVER IMAGE: A newly-discovered Leeuwenhoek microscope, Brian Ford, Hon FRMS A house clearance in East Anglia in December 2023 revealed this little microscope, which proved to have been hand-made by the Dutch pioneer Antony van Leeuwenhoek, some 350 years ago. Find out more by reading Professor Brian Ford’s fascinating article on p4.

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The Scanning Microscope and Leeuwenhoek’s Legacy Brian J Ford, Hon FRMS, Hon FLS

After some 350 years, newly discovered Leeuwenhoek microscopes are emerging. Two have now been subject to examination with the scanning electron microscope (SEM). Their maker, Antony van Leeuwenhoek, was the first microbiologist in history. He died 300 years ago, having shown us protozoa, bacteria, and spermatozoa for the first time. His work on plant anatomy was remarkably accurate, and he did much to dispel the dogma of spontaneous generation. I found his source of inspiration in Robert Hooke’s great work Micrographia, which was published in 1665 and was the talking point of London society when Leeuwenhoek made a visit to England the following year1.

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Hidden in the unnumbered pages of the preface

electron microscope (SEM) to examine them in

to Micrographia, and never cited by other scholars,

detail4. The bulk was left untouched, so that future

I found that Hooke had described in detail how

analysts could return to them for future study,

to make what we now call a Leeuwenhoek

uncontaminated by the modern world.

microscope2. Not only that, but the first specimens

There was a surge of public interest following the

Leeuwenhoek sent to the Royal Society (the

international publicity given to my discoveries, and

publishers of Micrographia) had been described by

an unknown Leeuwenhoek microscope was taken

Hooke, and in the same order3. After my discovery

to the Boerhaave Museum in Holland. Someone

of Leeuwenhoek’s original specimens, the Royal

had realised what it might be. This was not just a

Society allowed me to take small portions of each

fluke, for the interest continues – and others are

one to Cardiff University, where I am a Fellow, so

still coming to light to this day5. In December 2023

I could use the Cambridge Stereoscan scanning

yet another came to light. It was found during a

ISSUE 73 MARCH 2024

house clearance in East Anglia and was submitted to Christie’s auction house in London, who had sold one of the known Leeuwenhoek Microscopes in 2009 for close to half a million dollars6. This one didn’t make the mark; it cost the unknown Californian purchaser a mere £175,000.

Mr James Hyslop, head of science and natural history at Christie’s, spent many years at the Whipple Museum for the History of Science in Cambridge, and fully supported my proposal for scientific investigation, by examining the microscope in detail. He kindly agreed to bring it to the Cavendish Laboratory at Cambridge University for the day, so that I could scrutinise it under the SEM7.

The Quest for Authenticity This was the second Leeuwenhoek microscope I had analysed in this way. The first was a brass instrument found in the Netherlands in landfill mud dredged from a canal in Delft, Leeuwenhoek’s home town.

Museum

The reaction of the Boerhaave

(who

have

several

Leeuwenhoek

microscopes in their collections) was that this newly discovered brass microscope was one of their replicas that had been carelessly discarded.

Placed side-by-side with a known copy, they clearly have much in common8.

There was less doubt expressed over the silver instrument; nobody ever created replicas of Leeuwenhoek’s silver microscopes, due to the value of the materials. The way authenticity has always been established is based on provenance or personal judgement. For most of the surviving Leeuwenhoek microscopes there is limited documentation, and most lack a link back to Leeuwenhoek’s time. I set out to establish a protocol that would allow for objective authentication. The SEM allows us to discern how these little instruments were made, and that provides the evidence we need.

A house clearance in East Anglia in December 2023 revealed this little microscope which proved to be hand-made by the Dutch pioneer Leeuwenhoek some 350 years ago.The main screw was used to set the specimen at the desired height; a silver pin held it close to the single lens.

Low-power SEM is an unusual technique to apply to antique instrumentation. It combines images that have the astonishingly high resolution we expect

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This brass microscope was found during 2015 in mud from a canal in Delft, where Leeuwenhoek lived and worked. It was sold by a collector, along with a few coins from the Dutch East India Company, describing it as a ‘weird kind of drawing instrument’.

from electron microscopy, with the clarity we

methods of manufacture and assembly. The greatly

find when secondary electrons from a rasterised

increased resolution gives the unique insights we

scan are correlated to generate an image. The

need to establish authenticity.

reductionist aim with electron microscopy has long been to obtain images of greater magnification and higher resolution, though I have shown that submicron resolution in combination with low power magnification confers unforeseen benefits9.

Harnessing the SEM

dies, often oversized in order to imitate a handmade screw. Leeuwenhoek did not have such sophisticated equipment, and the threads of his screws were rolled. This method of manufacture displaces metal, rather than removing it. The SEM

Although large-chamber SEMs are available for

reveals the characteristic signs, notably the tendency

industrial applications, the specimen chamber

for the crest of the thread to be grooved. Detritus

of a conventional scanning electron microscope

within the root of the thread is further indicative of

imposes restrictions on the area that can be

its provenance.

scanned. Images captured in this case, using the Hitachi S-3400N SEM at the Cavendish Laboratory, University of Cambridge, measured ~1-2 mm so imaging a Leeuwenhoek instrument can involve the capture of ~100 separate files. Changes in focus or positioning can lead to minor perturbations in contrast, brightness, proportion or dimensionality, so assembling an image can require much manual

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Replica microscopes are always cut with industrial

The same procedure was used in creating the newly-discovered silver microscope. Evidence of production can be discerned on the specimen pin, which features a softly faceted profile due to its being hand-forged. A replica pin would have been turned on a lathe, and the SEM will immediately display the characteristic signs of tooling.

adjustment from frame to frame. Once completed,

Leeuwenhoek clearly wished his microscopes

the appearance of these diminutive instruments

to be functional, rather than perfectly finished.

is gratifyingly detailed and allows us to ascertain

Sometimes he decorated the main handle with

ISSUE 73 MARCH 2024

LEFT Replica Leeuwenhoek microscope of brass sold by the Boerhaave Museum, Leiden.

It is a faithful copy of an authenticated instrument in their collections.

CENTRE The brass microscope found by amateur detectorists in mud dredged from a Delft canal, claimed to be one of the Leiden replicas.The SEM proved this wrong.

RIGHT A silver Leeuwenhoek microscope, discovered during a house clearance in East Anglia, in December 2023. It has much in common with the Delft instrument.

three perforations, and the edge profile of each of his microscope body plates is always polished and rounded, whereas the brass sheet of the replicas is always cut with shears, and has a sharp, angular cross-section. Similarly, the perforated handles Leeuwenhoek produced are thin and seem neatly punched and polished, whereas the replica handles are thicker, and are conspicuously machined with a countersunk drill-bit. Even though Leeuwenhoek took pains to add this decorative feature, he made little attempt to ensure consistency of design – no two are the same – and bilateral symmetry was never his aim. One of the characteristics of his microscopes is their lightness. I found that the original brass microscope, for example, weighs a mere 6.24 g, whereas the mass of the Boerhaave

The recently discovered silver microscope under macrophotography at the Christie’s studio (left) compared with the finished SEM study (right). Details of the main screw just visible to the naked eye can be discerned with diagnostic clarity under the high-resolution SEM image.

pioneering biography in 187510, the instruments were mentioned in the past tense, as if all were then known. The microscopes in Leeuwenhoek's

Museum replica, with which some confused it,

possession that were left at his death were

weighs 14.71 g. Clearly Leeuwenhoek conserved his

auctioned in 1747, after his daughter Maria had

metals.

died. That was believed to be the last opportunity

Finding more Leeuwenhoek

for further unknown examples to emerge. Dobell’s master-work, published in 1932 to mark the

For more than two centuries it was accepted that

tercentenary of Leeuwenhoek’s birth, speaks of the

no further examples of Leeuwenhoek microscopes

remaining microscopes with no hint that any more

would ever be found. When Haaxman wrote his

might yet be found11.

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LEFT The stage assembly of the replica brass Leeuwenhoek microscope under the SEM discloses the lathe-turned specimen pin, die-cut screw thread, and prominent riveting.

RIGHT The specimen pin of Leeuwenhoek’s silver microscope, first revealed in December 2023, has a softer profile due to the malleability of the metal, and is clearly hand-made.

Since then, four previously unknown examples of

It proved to be the SEM that provided me with the

his microscopes have emerged, three of which had

means to establish a protocol by which these finds

passed through my hands. No scholar could have

could be objectively authenticated. It would now be

anticipated such a series of revelations. Clearly,

instructive to apply these findings to the remaining

unlikely as it seems, there may be others yet to

microscopes attributed to Leeuwenhoek, for the

emerge.

provenance of several is often weak, and their

LEFT Several of Leeuwenhoek’s brass microscopes feature his decorative pattern of three perforations on the main handle. A countersink drill bit has been used on this replica.

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CENTRE The recently-discovered authentic instrument from the Delft canal shows a hand-forged specimen pin, rolled screw thread, and nearinvisible rivet heads that are finely polished.

ISSUE 73 MARCH 2024

CENTRE Perforations in the handle of the authentic microscope were punched without a drill.The brass is also finer; a replica microscope weighs twice as much as a Leeuwenhoek original.

RIGHT Leeuwenhoek needed the construction of his microscopes to be functional, rather than precise.The silver handle was beaten to shape, with little attempt to make it symmetrical.

genuineness has been asserted by subjective opinion, rather than scientific facts. When I first learned of Antony van Leeuwenhoek when a schoolboy, there were nine known microscopes associated with his name.Today there are 13. But, were we to scrutinise them all with the guidance of the objective criteria that I have ascertained, several of those will prove to be copies or forgeries. Perhaps, at the end of all these proposed investigations, there may still be nine authentic Leeuwenhoek microscopes after all.

Acknowledgements The author is grateful to Professor Richard Langford at the Cavendish Labortory, and to JJ Rickard and Eric Tapley for technical assistance. The Royal

8: Brian J Ford, Recording Three Leeuwenhoek Microscopes, InFocus, Proceedings of the Royal Microscopical Society 40: 30-43, 2015.

9: Brian J Ford, New protocol for old microscopes, Laboratory News: 20-21, 2015 https://www.brianjford.com/w-1507-labnewsavl.pdf 10: PJ Haaxman Antony van Leeuwenhoek, de ontdekker der infusorian. Leiden: SC van Doesburgh, 1875.

11: Clifford Dobell, Antony van Leeuwenhoek and his ‘little animals’. London: John Bale, Sons & Danielsson, 1932.

Author

Email: brianjford@cardiff.ac.uk

Society generously funded part of this research,

Brian J Ford has studied Leeuwenhoek for over

published some of the results, and presented the

forty years. He discovered his original specimens

findings both at their Conversazione and Soirée.

hidden in the archives of the Royal Society,

References 1: Robert Hooke, Micrographia, or some

investigated how they were prepared, and was the first to take micrographs of a Leeuwenhoek section

through

an

original

microscope.

Physiological Descriptions of Minute Bodies, London, Royal Society: Martyn & Allestry, 1665.

Professor Ford has published many hundreds of

2: Brian J Ford, What were the Missing Leeuwenhoek Microscopes really Like?

Proceedings of the Royal Microscopical Society, 18 (2): 118-124, 1983.

on Leeuwenhoek alone, and many of his books

3: Brian J Ford, The van Leeuwenhoek Specimens, Notes & Records of the Royal Society 36 (1): 3759, 1981.

papers on microscopy, including 250 publications have been devoted to the microscope. He is a Fellow of Cardiff University, former Fellow of the Open University and Visiting Professor of Leicester University, and he is currently based at Cambridge.

4: Brian J Ford, Bacteria and Cells of Human Origin on van Leeuwenhoek’s Sections of 1674 [leading paper], Transactions of the American Microscopical Society, 101 (1): 1-9, 1982.

5: Dean Golemis, Editorial: Another Leeuwenhoek Microscope Comes to Light, The Microscope, 70 (3): ii, 2023 https://doi.org/10.59082/ KEAI2623 6: Gary Laughlin, Rare Leeuwenhoek bids for history, The Microscope 57 (1): ii, 2009. https://www.mccroneinstitute.org/uploads/ Editorial_57-1_2009-1477083872.pdf 7: Brian J Ford, The search for authenticity, Leeuwenhoek microscopes under the SEM, Microscopy and Analysis, 70: 15-17, 2024.

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Calendar We are very pleased to continue offering a range of ‘in-person’ and virtual events this year, in order to maximise accessibility and provide opportunities to those who might not otherwise be able to attend.

The following information was correct at the time of publication but could potentially be subject to change in the coming weeks. Please visit our event calendar at www.rms.org.uk for the latest updates.

If you have any questions about a booking you have already made for an event, or need any help or advice, please contact us at info@rms.org.uk

2024 March

5 Virtual International Microscopy Lecture Series - Dr Harald Hess, Online (Multi society event) 26 – 28 AFM & SPM 2024, Durham, UK

April 25

GW4 Cytomics 2024, Exeter, UK (RMS sponsored event)

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Laboratory-based X-ray Phase Contrast Imaging Workshop, London, UK (RMS-hosted event)

August 4–9

Strathclyde Optical Microscopy Course 2024,University of Strathclyde, UK (RMS-sponsored event)

September

2 – 6 Flow Cytometry Course 2024 York, UK

29 – 30 EBSD 2024, Glasgow, UK

June 4–7

elmi2024, Liverpool, UK (RMS-hosted event)

July

8 – 9 Light Microscopy Summer School 2024 York, UK 10 – 11 Getting the most from your Confocal Course 2024,York, UK

2025 March

26 – 28 flowcytometryUK 2025, Newcastle, UK

June / July

30 June – 3 July mmc2025: Microscience Microscopy Congress 2025, Manchester, UK

15 – 19 Electron Microscopy Summer School 2024 Leeds, UK

For further information on all these events, please visit our Event Calendar at www.rms.org.uk

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ISSUE 73 MARCH 2024

Featured RMS events EBSD 2024 29 - 30 April, Glasgow, UK Scientific organiser: Luke Daly, University of Glasgow The EBSD 2024 meeting will be held in person in Glasgow at the Mazumdar-Shaw Advanced Research Centre, on the 29 and 30 April 2024.

This Annual UK-based EBSD meeting is an excellent opportunity for the multidisciplinary EBSD community to meet and share the newest developments and applications of EBSD, and EBSD-related techniques, that are used to study materials across geoscience, materials science and engineering, and physical science. There are elmi2024 4 - 7 June 2024, Liverpool The European Light Microscopy Initiative was created in 2001 to establish a unique communication network between European scientists working in the field of light microscopy and the manufacturers of their equipment. Its aim is to promote the quickly developing field of light microscopy as a fundamental research tool for

also exciting emerging applications of EBSD to the biological sciences. Talks will likely include state-of-the-art developments in instrumentation and software, new techniques, as well as a variety of applications and uses of EBSD, transmission Kikuchi diffraction (TKD), electron channelling contrast imaging (ECCI), and related microscopy modalities.

As part of this series, we continue to be excited to hear from those who use these techniques to further our understanding of applied science and engineering challenges, as well as industrial, energy and environmentalchallenges (including the use of EBSD data in Industry 4.0 and for the energy transition).

the life sciences and to strengthen the channels of communication between researchers, core facilities and industry.

The annual meeting, which has been running for two decades at various venues across Europe, has an excellent reputation within the microscopy community, making this meeting a key event in the calendar of hundreds of scientists and developers.

The strength of this meeting lies in the mixture of scientific lectures on state-of-the-art, high-end microscopy combined with “hands-on” workshops and exhibition of the latest technology, organised by the leading companies in the field.

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Light Microscopy Summer School 2024 8 – 9 July, York, UK Scientific organiser: Peter O’Toole, University of York The Light Microscopy Summer School is a two day course held at the University of York covering the principles of light microscopy. Participants are also trained in practical issues surrounding light microscopy. After introductory presentations, the Getting the most from your Confocal Course 2024 10 – 11 July, York, UK Scientific organiser: Peter O’Toole, University of York This two-day, annual confocal course utilises many different sample types and fluorescent probes (DNA stains, classic antibody labels and fluorescent proteins) which are chosen to best demonstrate particular problems and techniques.

Focus is always on the techniques they enable Electron Microscopy Summer School 2024 15 – 19 July, Leeds, UK Scientific organisers: Louie Aspinall, Nicole Hondow, Rik Brydson; University of Leeds

ISSUE 73 MARCH 2024

course is taught predominantly through hands-on practical sessions. The course is suitable for both novices and more experienced users wanting to gain a greater understanding of the microscope and feedback every year is always fantastic. Students usually come from a range of backgrounds, within both research and commercial organisations. All benefited greatly from the course and left with increased understanding and skills. The course is immediately followed by a two-day, hands-on Confocal Course (see below). and the problems they generate, which will be applicable to any sample types. The two days consist of short tutorials followed by hands-on practice.

Day 1 takes participants through the basic principles of confocal microscopy and then trains them, through hands-on practice, how to configure and image multicolour, multidimensional samples using a confocal microscope.

Day 2 builds on the experience of Day 1 and enables participants to try FRAP and spectral profiling.

The Electron Microscopy Summer School aims to provide a basic training in both the theory and practice of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The course covers imaging, diffraction and chemical microanalysis as well as the highly important area of sample preparation.

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R E P O RT

Global BioImaging workshop Cuernavaca, México

“Facility management: Impact, facility operation, user training and image data” 21-24 Feb 2023

D

uring the week of 2124 February, 2023 Global BioImaging (GBI) and the Mexican National Laboratory for Advanced Microscopy (LNMA) organised a workshop titled “Facility management: Impact, facility operation, user training and image data” in Cuernavaca (México).

The workshop has gathered facility staff and bioimaging experts from Latin America, USA, Canada, Africa, Asia, Europe, and Australia. Several tens of countries were represented!

GBI

(https://globalbioimaging.org/)

is

“an

international network of imaging infrastructures and communities” that support training and job shadowing between facilities around the globe.

The LNMA (https://lnma.unam.mx/) is a network

of affiliated laboratories developing and offering advanced imaging techniques to academic and industrial partners across Mexico. It is supported by the distributed national university of Mexico (UNAM) and the CONACYT research council.

During the workshop, participants were taught how to measure the impact of facilities, identifying key performance indicators that align well with the values of the institution, and socio-economic indicators to evaluate the effects on the community, in terms of contribution to the overall education and technology development.

The intricate world of facility management was delineated, helping the participants to understand better how to manage microscopes, budget for the facility, offer efficient training, and deal with user expectations. The lessons on leadership and management held by Claire Brown, Phil Hockberger and many other trainers were invaluable.

(Left) Exchange of experience talk held by Rocco D’Antuono. (Middle) Pyramids in Xochicalco. (Right) In the company of Dr. Julián Mejía, postdoc working on the development of computational methods for super-resolution.

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ISSUE 73 MARCH 2024

Participants of the Global BioImaging workshop on Facility Management - Cuernavaca, México, 21-24 Feb 2023.

In between training sessions we had the opportunity

experience during the many group activities and

to listen to seminars held by local speakers who are

talks that a few of us were invited to do about how

working to improve the education system across

our facilities work.

the country.

After the workshop I could finally meet the

Particularly impressive is the effort of UNAM

postdoc that I helped recruit during one of our

university to implement ISO quality standards

collaborations with the research group led by Adán

across 3,000 research labs that host half a million

Guerrero at the LNMA. This is a sign that “what we

students in the country. This mission aims to

do always has an impact”, as we learnt during the

provide every student with the same teaching,

workshop.

guaranteeing the quality of research methods and

The workshop took place in Cuernavaca, state of

reliable standards for pharmaceutical research. The

Morelos, which is in the centre of the country and

quality management system has been adopted by

located close to many marvellous world heritage

33 faculties and institutes and relies on 150 internal

sites such as Xochicalco pyramids, the systems of

auditors and 500 academics specialised in quality

canals in Xochimilco and the world’s largest system

control (seminar held by Monica Gutierrez, UNAM).

of caverns that is the Cacahuamilpa national park.

A further initiative - the “Mexican Bioimaging

I would like to thank the Royal Microscopical

Workshops” - is aiming to reverse the concept of

Society and the Francis Crick Institute for allowing

centralised knowledge, by organising microscopy

me to attend this Global BioImaging workshop,

and image analysis workshops across the country,

which was very well organised by the local host

contributing with delocalised education to beat

Adan Guerrero (IBT, LNMA) with the support of all

social inequalities (Diego Delgado-Álvarez).

the wonderful members of the LNMA lab.

Personally, taking part in the workshop has been

Rocco D’Antuono FRMS

an enriching experience, allowing me to network

Principal Microscopist and Image Analyst

with many colleagues working in imaging facilities.

Crick Advanced Light Microscopy STP

I could get their perspective and learn from their

The Francis Crick Institute, London, UK

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2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT

Imaging the structural organisation of the LINC complex cage at the nuclear envelope using high resolution scanning electron microscopy Student: Lakshana Baheerathan Supervisor: Professor Martin Goldberg Project location: Durham University, Imaging Facility Lay Summary The cytoskeleton is anchored to the nucleus

On the other hand, the LINC complex could also

(specifically, the nuclear lamina) via the versatile

form a cage around the nucleus which attaches to

connectors collectively named as the LINC

the cytoskeleton. This implies, for instance, that

complex. LINC consists of inner nuclear membrane

mechano-sensing, via the cytoskeleton stretches

proteins (SUN1/2), which bind to the nuclear

and results in the tensioning of NE and connected

lamina and chromatin, and link, via their C-terminal

chromatin broadly, rather than in a localised area.

SUN domain in the nuclear envelope (NE) lumen,

As opposed to the point anchorage structure, this

to the C-terminal KASH domain of outer nuclear membrane nesprins. Interactions between the various SUN and KASH pairs provides a direct physical link between the cytoskeleton and nucleoskeleton and LINC plays a crucial role in the bio mechanical properties of the nucleus (such as stiffness, mechanotransduction, chromosome positioning). Consequently, it may have critical roles in controlling nuclear deformability and integrity during cell migration, with implications for metastasis and genome instability. Depending on the isoform of nesprins, they directly or indirectly link

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cytoskeleton which radiates out from the nucleus.

would greatly contribute to the biomechanical properties of the nucleus and influence its deformability during restricted migration.

Aim There is indirect evidence using immunofluorescence microscope and F-actin binding assays that suggest that nesprins could form similar structures at the NE. However, such a structure has not been directly imaged and this is the aim of the project.

Addressing the aim Nesprin 2 Giant molecules are 800kD in size

to cytoskeletal filaments. For example, Nesprin-1

with a distinct extended structure and previously

and -2 giant isoforms tether F-actin directly through

established extraction and fixation (aldehydes and

their N-terminal actin binding domain.Thus, nesprin-

tannic acid, OsO4) protocols, followed by critical

cytoskeleton complexes, LINC complex, form a 3-D

point drying and sputter coating with ultra-fine

filamentous network that covers the NE exterior.

platinum allows them to be imaged. However,

Current models portray the LINC complex as

the main challenge was to expose the nuclear

simple anchorage points at the nucleus for the

surface enough while still retaining the cytoskeletal

ISSUE 73 MARCH 2024

Figure 1. a) protein mound anchors with filaments spreading out across the ONM (mag x100k) using 530nm cryosection vehicle cells. b) Trimeric structure connecting the cytoskeletal architecture to the nuclear membrane. Nuclear baskets are also visible on the nucleoplasmic side of the INM (mag x100K); visualised from 1000nm cryosection vehicle cell.

attachments in order to image and interpret the

also tried different drying methods such as critical

hypothesised Nesprin 2 Giant network.

point drying and HMDS and found that the latter

Initially, we used a standard cytoskeletal SEM sample preparation on silicon chips followed by coating with 3nm of platinum and a simple dry-fracture process which exposes the nuclear surface to varying degrees. We optimised this protocol by testing

was much better for whole cell morphological visualisation. The optimum method to access the nuclear surface and reduce preparation artefacts was to cryo-section saponin-detergent extracted cells.

a variety of extraction detergents and incubation

Furthermore, we used a gelatin-based post-

periods and found that 0.75% saponin extraction

embedding immunogold labelling SEM protocol of

buffer with an incubation period of 20 minutes best

ultrathin thawed cryo-sections, in accordance with

exposed the layers of the nuclear membrane while

the Tokoyasu method. Well characterised (primary)

retaining cytoskeletal filamentous attachments.

antibodies to N- and C-terminal ends of Nesprin

However, upon fracturing with an adhesive we

2 Giants (and secondary labelling with 10-nm

found that the nuclei and often the entire cell is

colloidal gold) were used to elucidate doubts about

completely removed, leaving only cell remnants. We

the identity of structures that persistently appear

Figure 2.Work flow for immunogold labelling of 1000nm cryosectioned vehicle cell. (a) shows context of the region of interest; cytoskeleton and chromatin visible, yellow circle outlines perinuclear region where endoplasmic reticulum is located. Secondary image (b) and backscattered image (c) are taken simultaneously and then superimposed and tinted to reveal regions with protein of interest (d). All images have a magnification of 100K.

17

Figure 3. a) All 3 layers of the membrane; outer and inner nuclear membrane (ONM and INM) and (putative) nuclear lamina (NL) is visible.

Nuclear membrane in a 1000nm cryosection.White arrow points to a nuclear basket on the nucleoplasmic side of the INM.Yellow point shows cytoskeletal attachment and how the structure is continuous and passes through the nuclear envelope and is connected to the nucleoplasm and related structures on the other side. (B) Nuclear membrane in another 1000nm cryosection. Here white arrows point to regions where a continuous structure that mimics a tunnel passes through the envelope and is connected to the lamina and nucleoplasm.

on the outer nuclear surface. Labelled sections

at the nuclear envelope. However, optimisation of

were coated with 3nm chromium and were

the protocol involving improvements in antibody

examined in the SEM. Images from secondary and

selection line and dilution must be made for more

backscattered images are acquired simultaneously

specific and reliable binding.

and are superimposed to show the location of gold (labelling the protein of interest) on the secondary image.

parallel and highly organised network of filaments (with a diameter ranging 10-15nm) that line the nucleoplasmic side of the inner nuclear membrane.

Results Our results provide preliminary evidence that combines both the point anchorage and cage structure model of the LINC complex at the nuclear envelope. Images depict a large protein or group of proteins that are regularly spaced at intervals ranging from 150 – 200nm. I hypothesise that these protein mounds act as an anchor that connects the cytoplasmic cytoskeletal architecture directly to the outer nuclear membrane (ONM). Spreading from this point are filaments with diameters comparable to actin and intermediate filaments, but short, finer fibres have also been visualised. I believe that the protein anchor could act as a potential nucleation site that allows the extension of these filaments.The filaments spread over the surface of the ONE and appear to connect to neighbouring protein mounds.

18

Imaging at higher magnification also reveals a

Based on previous literature, I hypothesise that this network is the nuclear lamina. The network can be described as a sort of railroad that lies directly on the inner nuclear membrane and imaging at higher magnification revealed that heterochromatin and nucleosomes are associated or somehow bound in a way to this network. This structure is similar to those previously found in research associated with identifying a nucleoskeleton. Imaging also revealed that the filaments that spread over the surface of the ONE forming a cage make direct contact with the ONE and pass through the nuclear envelope and embed in the inner nuclear membrane.

What I learned I learnt a vast array of sample preparation techniques including cell culture, cryosectioning,

Immunogold labelling revealed that the hypothesised

glass knives making and critical point drying. While

protein anchor are (or contain) Nesprin 2 Giants.

anxious at first, I became much more confident in

Immunogold

that

my abilities and understood the importance of not

pAbK1 Nesprin-2-specific antibodies recognise

putting too much pressure on the experiments

both nuclear and cytoplasmic Nesprin-2 proteins

themselves. Working primarily with scanning

SEM

ISSUE 73 MARCH 2024

examination

revealed

electron microscopy, I gained a deeper appreciation for the physics underlying the principle of the SEM and learnt how to adapt the sample preparation and imaging techniques. To balance the resolution and magnification with sample contamination I tried out different sectioning and fracturing techniques and used various accelerating voltages.

I really enjoyed the freedom of designing and carrying out experiments independently and learning and adapting as I went along. I also had an opportunity to present and discuss my ideas during different stages of the studentship, and this allowed me to critically analyse and produce original ideas that directed and fuelled subsequent stages of the project.

Lakshana Baheerathan.

This is the first independent project that I have conducted and it has given me both technical and

How this project affected my long term goals?

transferable skills that I hope to use as I progress in

Prior to this summer, my research interest tended

Following this summer, I am conducting a research

to mostly lie around biochemical analysis, but

project focusing on the biosynthesis of collagen

this summer studentship has given me the time

and will be using confocal and immunofluorescence

and experience to confirm my interest in cell

microscopy and live cell imaging. Additionally, I am

biology research. While I have always appreciated

keen on undertaking a master course targeted

the quantitative side of research, the freedom to

on understanding molecular and cellular signalling

critically interpret visual data and be creative with

processes and aim to incorporate microscopy as a

it is incredibly exciting as it fuels further research.

significant component of the project.

the world of research.

Contacting the Royal Microscopical Society The offices of the Royal Microscopical Society are at: 37/38 St Clements, Oxford, OX4 1AJ, UK Tel: +44 (0) 1865 254760 For general enquiries email info@rms.org.uk For information about meetings and courses email events@rms.org.uk For membership enquiries email membership@rms.org.uk

www.rms.org.uk

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2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT

Mechanical Meniscus Correction in Phase Contrast Microscopy Student: Mohamed Y. Ghali

Supervisors: Kevin F. Webb, Optics & Photonics Research Group; Steve Greedy, George Green; Institute for Electromagnetics Research Project Location: Electrical & Electronic Engineering, University of Nottingham Imaging living cell samples is challenging due to the meniscus formed by immersing liquid at container walls. The meniscus distorts the passage of illumination, creating artefacts in transmitted light microscopy. This is especially a problem with phase contrast microscopy1, where the illuminating annulus must match the phase ring to produce phase contrast (as in Figure 1).

The effect of this meniscus distortion can be

axis robot arm, Figure 2) to physically reposition the

captured with an image of the microscope back

illuminated annulus in space, utilising the condenser-

focal plane (Figure 1, insets), which quantifies the

free illumination regime developed by Webb 2.

misalignment between the phase ring (dark ring) and the illuminating annulus (bright ring). This mismatch can affect the XY-position, diameter, and circularity of the illumination (inset, Figure 1B).

The approach taken is described in the flowchart in Figure 3, where image processing (OpenCV) was used to locate and measure dark and bright ellipses in images of the back focal plane. Geometric

This project aimed to physically correct this

parameters describing the distortion (Figure 4)

misalignment using a mechanical actuator (multi-

were determined in real time, in order to guide the

Figure 1. A). Phase contrast image, showing the benefit of correct alignment of a phase contrast microscope (inset). B) Meniscus-misaligned phase contrast image, showing the effect of the meniscus distortion to the illumination annulus (inset) on image contrast.

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ISSUE 73 MARCH 2024

of the image processing algorithm, showing ring misalignment caused by a water droplet. Figure 4b shows the correction factors required to superimpose the illumination and phase rings.These corrections were sent via UART data interlink to physically reposition (in XYZ) and incline the ring in order to achieve the required correction via the robot arm (Figure 1). The project will now be furthered through my capstone final year individual BEng project in Electrical & Electronic Engineering.

Prior to this project, I had used microscopes a few times during biology experiments in school, where usage comprised blindly following instructions with very little understanding. During this project, my knowledge about optics and microscopy was vastly expanded. I gained familiarity with various Figure 2. Development rig, showing multi-axis robot arm positioning LED ring illumination above a multi-well plate under microscopic observation.

motion of a multi-axis robot arm (WidowX-200, Trossen Robotics, IL) controlled by an OpenCM9.04 microcontroller3, which held a 100mm diameter white COB LED ring (eBay). This functioned as the condenser-free phase contrast illuminator for a Leitz Diavert inverted microscope.

A Raspberry Pi 4B single-board computer, equipped with v2.1 Pi camera, was focussed on the objective

microscopical tools for phase contrast microscopy in particular, including phase telescopes and the relationships between image and back focal planes.

Further skills which are vital to any EEE engineer were also learned/strengthened, including 3D modelling/prototyping, image processing, UART communications,

and

3D

kinematic

control

architectures.

Whilst completing the final year of my BEng degree, my aim is now to look for support for a PhD programme (ideally in the UK), with the

back focal plane using a phase telescope. OpenCV

goal of an academic or research career. Prior to

used a thresholded binary image of the back focal

this summer studentship, my PhD ideas revolved

plane to fit an ellipse to the illuminating ring pixels.

around any subjects utilising IoT technology and/

This was then compared to the hardwired phase

or autonomous vehicles. This summer project

ring within the objective to determine the relevant

catalysed a new interest in further specialisation

corrections

and

on applications involving optics and/or microscopy,

inclination) to correct the size, shape, and position

(XY-translation,

eccentricity,

perhaps using information extracted through

of the illuminating ring to again superimpose

microscopy

with the phase ring. Figure 4 shows the output

(temperature, humidity etc.) in an incubator, or to

to

control

different

parameters

Figure 3. Schema of a system mechanically correction meniscus artefact by physically repositioning a ring using a robot arm.

21

2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT References 1.

Hofmeister, A., Thalhammer, G., Ritsch-Marte, M., & Jesacher, A. “Adaptive illumination for optimal image quality in phase contrast microscopy”. Optics Communications, 459, 124972, 2020, doi: doi:10.1016/j. optcom.2019.124972.

2.

Webb, K.F., “Condenser-free contrast methods for transmitted-light microscopy”. Journal

Figure 4. Back focal plane image analysis showing A) Misaligned illumination ring, fitted with an ellipse (red), superimposed but displaced from the phase ring (green). B) Relevant correction parameters output from the algorithm (in pixels).

guide autonomous vehicles transporting biological cargo to optimise their travel (slow down; take a break; open windows etc.) to cater for their biological cargo.

Mohamed Ghali.

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ISSUE 73 MARCH 2024

of Microscopy, 257: 8–22, 2015, doi:10.1111/ jmi.12181.

3.

‘WidowX 200’, WidowX 200 Robot Arm. Accessed: Sep. 26, 2023. [Online]. Available: https://www.trossenrobotics.com/widowx200-robot-arm.aspx

Special Issue Imaging Cell Architecture and Dynamics Guest Editors: Lucy Collinson and Guillaume Jacquemet Extended deadline: 29 March 2024

Call for papers Find out more at: journals.biologists.com/jcs/pages/imaging Image credit: Guillaume Jacquemet. See article by J. W. Pylvänäinen et al. (jcs260728).

2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT

Evaluating metrics used in deep learning for microscopy Name of Student: Miriam Czech Supervisor: Susan Cox, Siân Culley Project location: Randall Centre for Cell & Molecular Biophysics, King’s College London Summary

evaluation of image restoration networks used for

Fluorescence microscopy is an essential tool in

enhancing the quality of microscopy images.

cell biology as it allows scientists to look at what is inside of a cell and easily see the organisation of

Methods

organelle by labelling different organelles selectively.

To achieve the aim, the two architectures were

One of this technique’s major limitations is that the

examined against images with different amounts of

light needed to image a live sample can also damage

structure, and different peak signal values.

it, if applied at too high an intensity or for too long.

After training a network, its predictions were

This leaves the researcher needing to choose between a limited duration of imaging or image artefacts introduced by photo-damage. A possible solution is imaging at a low laser intensity and later computationally restoring acquired images (Weigert et al., 2018). My project focused on investigating two state-of-the art deep learning methods used for image restoration in microscopy. The two learning networks were trained on synthetic images. Their performance was evaluated against parameters of the laser intensity of imaging and the structural content density in terms of four restoration quality

of the conventionally used metrics - Normalised Root Mean Squared Error (NRMSE), Peak Signal to Noise Ratio (PSNR), Structural Similarity Index (SSIM) and Multi-Scale Structural Similarity Index (MS-SSIM). The networks were also benchmarked against the simplest possible denoising technique of a Gaussian blur. Furthermore, the analysis was complemented by looking at the pixel values’ histograms and qualitative inspection of chosen pairs of images with inspection ImageJ tools.

assessment metrics that are conventionally used

To easily obtain an adequate volume of training and

for assessing denoising methods. The performance

evaluation datasets with a wide, fine-grained, and

of the networks at background and foreground

systematically defined spectrum of parameters’

prediction was also compared. Finally, a comparison

values, synthetic images were generated. For each

to the simplest denoising technique of applying a

parameter, three distinct values were chosen

Gaussian blur was made.

for training and 20, within the same range, for

Aim The project’s aim was to better understand two popular denoising networks, CARE (Weigert et al.,

24

compared with their source ground truth in terms

evaluation. Poisson noise was applied to simulate the sort of noise expected in low power intensity microscopy images.

2018) and Noise2Void (Krull et al., 2019) and to

Findings

test the appropriateness of four quality assessment

Surprisingly, CARE did not outperform the

metrics conventionally used in training and

technique of Gaussian blur (GB) for most parameter

ISSUE 73 MARCH 2024

Figure 1. Investigating the sensitivity of the metrics (NRMSE, PSNR, SSIM, MS-SSIM) to the number of blobs by which images compared differ.

combinations. Its performance was especially poor for low structural content density and low signal.

It was hypothesised this stems from an intensity correction the algorithm applies to deal with pixels of very low and high intensity. These are returned by the camera because of a Gaussian laser beam profile - an artefact caused by most common lasers.

To investigate this, a large background Gaussian was added leading to better performance for all parameter combinations according to NRMSE, and improving most combinations according to other

structural content combinations.

An N2V network was trained with images with a Gaussian background. According to NRMSE, PSNR and SSIM, on average it did not outperform GB for only a minority of parameter combinations with a low structural content and low signal peak value.

In terms of MS-SSIM, N2V outperformed GB in most cases, failing for high density, low signal values combinations. Its restoration improved images for all combinations in terms of NRMSE, PSNR and MS-

metrics. To understand the impact of altering the

SSIM and SSIM.

structure on the metrics, the impact of changing

The background was predicted less successfully than

the number of blobs in the model was evaluated for

the foreground by the N2V network. However, the

each metric (see Figure 1).

maximum discrepancy between the two was much

The performance of the network architecture

smaller than for CARE. In terms of NRMSE, N2V

proved to be influenced by the diversity in the

seems to have a more consistent performance

training data. Another network was trained on

across different levels of structural content and

a narrower spectrum of signal values and the

a more balanced one between background and

performance was improved, except for low

foreground, whereas CARE seems to do better than

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2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT N2V with high structural content and high signal

premises of neural networks in more detail and

value.

motivated me to dive deeper into how neural

From a visual inspection we found that the errors in the background are unlikely to be spotted by eye.

Since in microscopy the background pixels usually dominate, the overall metric scores get dominated by the background error. Hence, the overall image metric scores are possibly not reflective of the

network architectures are designed. I also got very excited about developing further my ability to use the PyTorch framework. I had not known much about computer vision and the internship made me excited for some potential related projects in the future.

actual quality of restoration if the accuracy of the

I was also able to witness the day-to-day life of a lab

foreground is the primary consideration. Thus, this

researcher and was especially captivated with how

study shows the need to better understand how

much autonomy there is in the work and to what

the restoration technology underpinning evaluation

a large extent it seems to be dictated by where

metrics works for fluorescence microscopy images.

their curiosity takes them. My internship experience

Learnings I really enjoyed learning to use the PyTorch

added to arguments in favour of pursuing a PhD someday.

framework and figuring out how to do operations

Sources:

on tensors to achieve high code efficiency when

1. Krull, A., Buchholz,T.-O. and Jug, F. (2019)

operating on large volumes of data. Discussing with my supervisors the possible explanations for the results we would get was extremely developing and exciting. I also loved going to my host lab’s weekly lab meetings and listening about what lab members were working on at the time.

‘Noise2void - learning denoising from single noisy images’, 2019 IEEE/CVF Conference on Computer Vision and Pattern Recognition (CVPR) [Preprint]. doi:10.1109/cvpr.2019.00223.

2. Weigert, M. et al. (2018) Content-aware image restoration: Pushing the limits of fluorescence

I learned about the basic terms relating to neural

microscopy, Nature News.Available at: https://

network training and performance evaluation,

www.nature.com/articles/s41592-018-0216-7

how to use TensorFlow’s TensorBoard to help

(Accessed: 16 October 2023).

with improving the quality of training and ways of identifying whether a network is undertrained or overtrained. Moreover, I learned the basic premises of neural networks such as the gradient descent algorithm, the fundamental structural elements of a neural network and about the U-net convolutional neural network architecture in more detail.

Participating in the weekly lab meetings and the discussions with my supervisors taught me about the current limitations of microscopy as physical science and the applicability of computational techniques as possible ways forward and types of image restoration assessment methods.

Impact on long-term goals The project allowed me to explore the mathematical

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Miriam Czech.

Visualising Bruton’s Tyrosine Kinase Inhibitors using Stimulated Raman Scattering Microscopy Student: Hio U Lao Kathleen Supervisor: Dr William Tipping, Professor Duncan Graham Where the project took place: University of Strathclyde, Glasgow Supported by an RMS Summer Studentship award, I spent time in the lab of Professor Duncan Graham at the University of Strathclyde to study the uptake and distribution of Bruton’s tyrosine kinase (Btk) inhibitors, ibrutinib, acalabrutinib and ibrutinib-yne using a combination of Raman and stimulated Raman scattering microscopy. Raman imaging techniques are beginning to shed new insight into drug localisation and drugcell interactions without the use of exogenous tags and labels.

We sought to detect Btk inhibitors at their site of action in the cellular environment to detect differences in uptake concentration and localisation in a panel of cell lines expressing different levels of Btk. We also investigated the impact of Btk inhibition on cellular lipid metabolism.

Lab Summary:

(second generation) are approved Btk inhibitors,

Bruton’s tyrosine kinase (BTK) was known to

whilst with ibrutinib-yne contains a phenylacetylene

be highly expressed in B cells, and some breast

motif that provides access for click chemistry

and prostate cancer cell models. It is a crucial

applications. We sought to determine the uptake

mediator in cellular function and is involved in B cell

and localisation of these molecules across the panel

proliferation. Here, we used Raman microscopy and

of cell lines together with detection of ibrutinib-

stimulated Raman scattering (SRS) microscopy to

yne using the alkyne tag as a marker in the Raman

visualise the cellular uptake of Btk inhibitors (Btki)

spectrum.

in cancerous cell lines. We selected cell lines which express low levels of Btk including the ovarian cancer cell model HeLa and the hepatocellular carcinoma HepG2 cells, whilst the prostate cancer model PC3 and chronic myeloid leukaemia cell line K562 were used because these cell lines express moderate levels of Btk. Three Btk inhibitors were selected: ibrutinib (first generation) and acalbrutinib

Various concentrations of Btki were treated on cell lines for 4hr, 18hr and 24hr. However, no alkyne peak was observed in both Raman mapping and SRS results of cell lines that express low levels of Btk (HeLa and HepG2). Lipid droplet analysis showed that in the prostate cancer cells, PC3 resulted in higher percentage of lipid droplets

27

2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT than compared to the prostate healthy, PNT2

structure of ibrutinib-yne allowed label-free imaging

cells. Following treatment with Btk inhibitors, we

of the drug uptake using Raman microscopy and

detected a reduction in lipid droplets after drug

SRS microscopy.

treatment which we predicted was related to the cytotoxicity of drug effects. To test this, cell viability

Methodology:

assays were performed using AlamarBlue which

Cell culture

showed Ibrutinib induced a higher cytotoxicity effect compared to Acalabrutinib, which correlated to a greater lipid droplet reduction in these cells. In K562 cells, we were able to detect the localisation of ibrutinib-yne using the Raman signal of the alkyne group at 2107 cm-1 indicative of the alkyne vibration. The signal was concentrated in small punctate in the cell cytoplasm which we propose could be the lysosome, which is an organelle used for the degradation of waste products in the cell.

Human cancerous cell lines (HeLa, HepG2, PC3 and K562) and normal cell line PNT2 were split and cultured in DMEM and RPMI respectively. Cells were treated with concentrations of 2-50 μM for a range of timepoints (1hr, 4hr, 18hr, 24hr) and were incubated at 37°C with 5% of CO2. Overnight serum starvation (-FBS) was also performed in all cell lines.

AlamarBlueTM assay Cells were cultured and incubated overnight with

Studies are currently ongoing to identify the nature

excess PBS filled in the outer wells. HeLa cell lines

of this interaction using a multiphoton Raman and

were treated with Ib and Acal in concentrations

fluorescence imaging approach.

ranging from 100 μM to 0.03 μM for 72hr. PC3 and

In conclusion, although low Btk-expressed cells were

K562 cell lines were treated with Ib, Acal and Ibyne

not able to be screened using Raman microscopy, our further approach is to work on screening high BTK expression cells using Ibrutinib-yne.

with the same concentration range and time. 10 μl of Alamar Blue was used in staining for analysis following 4hr incubation in each well.

Raman imaging

Aim: We focused on treatments using Btk inhibitors Ibrutinib (Ib), Ibrutinib-yne (Ibyne) and Acalabrutinib (Acal) on cell lines that consist of relatively low Btk expression (HeLa and HepG2) together with cell lines that expressed moderate Btk levels (PC3 and K562). The presence of alkyne moiety within the -1

1440/1656 cm cellular lipids

Raman spectra and mapping were obtained at 532 nm using the Renishaw microscope, with a 60x objective lens, 1 μm step size in x and y coordinates, 5s acquisition time, 36 mW laser power and 1500cm-1 for the spectral centre.

For SRS microscopy images were captured at

1340/1656 cm DNA

-1

Figure 1. Ratiometric Raman imaging of fixed HeLa cell to detect the cellular lipid, DNA and protein content.

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ISSUE 73 MARCH 2024

-1

1003/1656 cm cellular proteins

-1

2930 cm Proteins

2851 cm

-1

Lipids

PNT2

PC3

Figure 2. SRS imaging of the cellular proteins and lipids in PNT2 cells and PC3 cells treated with ibrutinib (5 μM, 4 h).The protein signal was detected using the CH3 stretch at 2930 cm-1 and the lipid signal was detected using the 2851 cm-1 band associated with the CH2 stretch. Scale bars: 10 μm.

2930cm-1 aa 2851cm-1 regions indicative of cellular

believe this may be due to toxicity causing the cells

proteins and lipids. Alkyne images were acquired

to use stored lipids for energy demands. Additional

at 2107 cm-1. To do so, the pump and Stokes laser

Raman mapping was performed on CaF2 plates

powers were set to 0.15 W and 0.3 W respectively.

which showed a reduced spectral background. We

The image acquisition parameters were as follows:

then elected to use SRS microscopy to visualise

410 v gain, 10Hz acquisition rate, across a 512 ×

the prostate cell lines due to the faster image

512 frame.

acquisition rates for SRS microscopy compared to

Results and Discussion:

Raman microscopy. SRS microscopy showed that the PC3 cells had more lipid droplets than the PNT2

Raman mapping was first performed in HeLa

prostate healthy cells when treated with ibrutinib (5

cells which showed that label-free ratiometric

μM, 4 h) because an increase in the signal at 2851

imaging of the protein, lipid and DNA content was

cm-1 (CH2 stretch, Figure 2) was observed.

possible (Figure 1). The cellular lipid content was detected at 1440 cm-1/1656 cm-1 (CH2/amide-1), nucleic acid contents at 1340 cm-1/1656 cm-1 (C-N mode/amide-1), and protein content using the phenylalanine stretch at 1003 cm-1/ 1656 cm-1 (phe/ amide-1). The ratiometric imaging revealed a labelfree method for cellular visualisation based on the vibrational spectrum.

K562 cells which express high levels of BtK were treated with ibrutinib-yne (5 μM, 4 h) and SRS revealed an alkyne signal was detected at 2107 cm-1 indicative of the alkyne stretching frequency (Figure 3). The alkyne signal was not detected in K562 cells treated with ibrutinib (5 μM, 4 h) as expected.

However, in cells that express low levels of Btk, we found that Raman microscopy might not be sensitive

In cells treated with Btk inhibitors, a reduction

enough for detecting alkyne signals following

in cellular lipids was observed at 1440 cm-1. We

treatment with ibrutinib-yne. Future studies will

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2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT

Figure 3. K562 cells treated with ibrutinib-yne (top) and ibrutinib (bottom) at concentration of 5 μM for 4 h. SRS images acquired of the cellular proteins, lipids and alkyne (ibrutinib-yne). Data acquired with A. Merchant (U. Strathclyde). Scale bars: 10 μm.

investigate the quantification and localisation of

and professors across Scotland. In conclusion, the

ibrutinib-yne in real-time to identify mechanisms of

experiences and skills I have acquired throughout

drug uptake and retention.

this studentship have provided me with the

Reflection:

motivation to work more independently, with great efficiency and attention to detail in my upcoming

Throughout the studentship, the research project

industrial placement. I am very grateful to the RMS

has been more challenging and inspiring than I

and the department of Chemistry for this valuable

initially anticipated. Engaging in the planning and

opportunity.

conducting of experiments has helped me to further develop a more flexible and comprehensive mindset. Through working both independently and collaboratively, I have not only strengthened my practical skills and reinforced my knowledge, but I have also learnt how to work and communicate efficiently with others. I particularly enjoyed the practical side of this project, from learning about cell splitting and culturing to treatments and operation of the Raman and SRS equipment. It has been an incredibly fascinating experience for me thus far. Furthermore, I was also delighted to have the opportunity to attend an imaging conference, where I had the chance to understand some of the cutting-edge research being carried out by students

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Kathleen performing a spontaneous Raman mapping experiment of a HeLa cell.

Investigating BRCA2 and MCAK Interactions with a Microtubule Depolymerisation Assay Student: Rebecca Harry, University of Oxford Supervisor: Professor Claire Friel, University of Nottingham Project Location: University of Nottingham, School of Life Sciences, Friel Lab Lay Summary

Kinesins are biological motor proteins that

Eukaryotic cells have a cytoskeletal structure which

work in an ATP-dependent manner, working in

helps to maintain shape and organise components

cells to transport molecules along microtubules.

of the cell. The cytoskeleton provides mechanical

The kinesin-13 family drive depolymerisation

support that enables cells to carry out essential

(breakdown) of the microtubules, contributing to

functions like division and movement. The key

the regulation of these fibres in the cell. In this

components of the cytoskeleton are microtubules,

project, I generated microtubules in vitro with

actin filaments, and intermediate filaments. This

rhodamine labelled tubulin and then followed

project explored how microtubules are built up and

their depolymerisation when MCAK (a kinesin-13

broken down dynamically inside eukaryotic cells,

family protein) was added. This data was then

contributing to the constantly changing cellular

analysed to calculate the rate at which the

environment and helping with cell division.

microtubules were depolymerised. This allowed

Figure 1. A box plot showing the results for the MCAK depolymerisation assays we carried out in the presence and absence of the 2 BRCA2 peptides Peptide 1 = phosphorylated, Peptide 3 = non-phosphorylated. The phosphorylated form is the form shown to bind to MCAK.

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2 0 2 3 S U M M E R S T U D E N T S H I P R E P O RT

Figure 2. Kymograph showing the effects of peptide 1 and peptide 3 on the ability of MCAK to induce depolymerisation in microtubules.

comparison between MCAK alone and MCAK in

depolymerisation assay to help gain understanding

the presence of various fragments of BRCA2, that

of how interactions that have been detected

have been shown to bind to MCAK, on the rate

between MCAK and BRCA2 affect the microtubule

of microtubule depolymerisation. This work allows

depolymerising

us to understand more about a recently discovered

possible therapeutic implications due to the role

interaction between MCAK and BRCA2 which

of microtubules in cell division and BRCA2’s well

could have therapeutic importance due to the role

known role as an oncogene.

of microtubules in cell division.

Project Aim The aim of this project was to conduct a

activity

of

MCAK. This

Addressing the Project Aim Using a depolymerisation assay allowed us to follow the process of microtubule depolymerisation by

Figure 3. Shows how microtubules were observed through the microscope prior to individual ones being selected for detailed dynamic analysis.

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has

MCAK. To set this up, microtubules were grown

I had not previously thought of microscopy as an

in vitro with rhodamine labelled tubulin and then

essential technique for in vitro protein study, so I

imaged using a high-resolution light microscope. In

was appreciative of the diversity in which it can be

order to conduct a depolymerisation assay, channels

used to study biological processes.

were engineered between two coverslips, and the allowed us to fix microtubules in the channels so

How has this project affected your long-term goals?

that they could be imaged in the same position

This project was my first real experience with

throughout the experiment. Next, MCAK was

working in a lab on a set project, due to various

added to channel at a set timepoint and then the

limitations that were in place due to Covid-19

process of depolymerisation captured via timelapse

earlier in my degree. I enjoyed working in a lab,

capture. Once a baseline rate for MCAK activity

with the opportunity to work independently but

had been established, we could then test the effect

still with the support of my supervisor, Professor

of each of the BRCA2 peptides on the activity of

Claire Friel. After completing this project, I will be

MCAK.

researching PhD programmes going forward, as well

channel then filled with anti-tubulin antibodies. This

as completing my Masters this coming year. Moving

What did you find out?

forward, I will continue to view microscopy as a

From completing depolymerisation assays with

key tool for the biochemist, useful in both in vitro

1) MCAK, 2) MCAK plus a BRCA2 peptide,

and in vivo processes, as well as static and dynamic

and 3) MCAK plus a phosphorylated BRCA2

situations. I will be incorporating microscopy into

peptide, we observed no significant effect of

my experiments investigating X chromosome

either BRCA2 peptide on the ability of MCAK to

silencing, using immunofluorescence to examine

depolymerise microtubules (Figure 1 and Figure 2).

efficiency of the process under different conditions.

These data indicate that binding of these BRCA2

I am extremely grateful to the Royal Microscopical

peptides to MCAK does not impact microtubule

Society for enabling me to complete this valuable

depolymerisation

experience and supporting my interest in the Friel

and

that

we

should

look

elsewhere for the functional impact of the observed

group’s work.

interaction between BRCA2 and MCAK.

What did you learn from participating in this project?

During this project I learnt how to grow microtubules in the lab so that they could be visualised and analysed via microscopy. I also learnt the importance of kinesins in depolymerisation in addition to their more traditional role as transporters along microtubules, moving key components around the cell. I also learnt more about how microscopy can be essential for tracking processes in the cell over a time period as well as for single frames which visualise just one moment. In particular, I enjoyed the process of starting from tubulin subunits and building up to a working assay with microtubules that could then be used to assess protein activity.

Rebecca Harry.

33

Missing Atom This image, which was shortlisted in the AFM & SPM category of the 2023 RMS Scientific Imaging Competition, shows atomically resolved crystal structure of MoS2 surface - clearly identifying a missing sulfur atom.This image was obtained via conductive atomic force microscopy under ambient conditions.

Image credit: Saima Aktar Sumaiya, Mehmet Baykara (University of California, Merced; Columbia University).

Bacterial skin Also shortlisted in the AFM & SPM category of last year’s imaging competition was this 3D height visualisation of the internal peptidoglycan structure of a Staphylococcus aureus mutant. It shows individual glycan strands, taken with AFM in liquid.The colour scales correspond to the height data (lower – green; higher - blue).

Image credit: Dr Laia Pasquina Lemonche (University of Sheffield).

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ISSUE 73 MARCH 2024

silver ink on newspaper Shortlisted in the Electron Microscopy (Physical Sciences) category of the RMS Scientific Imaging Competition, this is a coloured scanning electron micrograph of particles from a silver roller ball ink pen in their carrier (pink) on newspaper.

Image credit: Steve Gschmeissner.

Sub-units Shortlisted in the Electron Microscopy (Physical Sciences) category of the RMS Scientific Imaging Competition, this is an image from thin films of manganese dioxide, developed for confection of flexible polimeric structures.The acquisition was performed by scanning electron microscopy (JEOL- JSM IT500 HR) in the Multi-user Center for Analysis of Biomedical Phenomena (CMABio-UEA) located in the middle of the Amazon rainforest, in the north of Brazil.

Image credit: Jessica Araujo Marques and Jander Matos Guimarães (Universidade do Estado do Amazonas).

35

Delegates hit the dance floor as a band plays during elmi2018 in Dublin – the last time the RMS hosted an ‘in-person’ elmi.

Register now for elmi2024!

The RMS is very proud to be hosting elmi2024, taking place in Liverpool, UK, from 4 - 7 June.

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ISSUE 73 MARCH 2024

With a blockbuster scientific programme, worldclass exhibition, ‘hands-on’ workshops and more, the European Light Microscopy Initiative (elmi) has become regarded as an essential event for the Light Microscopy community.

‘Early bird’ booking rates are available until 9 April providing a discount of more than 20% on the cost of a standard ticket, and 30% off for students.

Find out more about registration rates and book now! the iconic city, on the King’s Dock.

The venue has great transport links as well as being walking distance from a large number of hotels, restaurants, shops and bars.

Liverpool is one of the UK's best loved cities with

Unique communication network

a rich, cultural heritage, a wide range of attractions and buzzing nightlife.

With the conference venue just a short walk from

The European Light Microscopy Initiative (elmi) was

the bustling waterfront and commercial centre,

created in 2001 to establish a unique communication

there is no shortage of things to see and do during

network between European scientists working in

your stay.

the field of light microscopy and the manufacturers of their equipment. Its aim is to promote the quickly developing field of light microscopy as a

What you can expect at elmi2024:

fundamental research tool for the life sciences

• A blockbuster meeting programme covering

and to strengthen the channels of communication

all the latest techniques, applications and

between researchers, core facilities and industry.

technology. Topics include: New Technologies,

The event has been running for two decades at

Imaging Across Scales, Super-resolution and

various venues across Europe, and has an excellent

Nanoscale Imaging, The AI Revolution, The

reputation within the microscopy community,

Science of Tomorrow Today, and Multimodal

making it a key fixture in the calendar of hundreds

Imaging.

of scientists and developers.

The strength of this meeting lies in the mixture of scientific lectures on state-of-the-art, high-end microscopy, combined with ‘hands-on’ workshops and an exhibition of the latest technology, organised by the leading companies in the field.

Great venue in the heart of Liverpool elmi2024 is being held at ACC Liverpool, a purposebuilt arena and convention centre in the heart of

• A wide range of companies showcasing their latest technology and running workshops timetabled outside of the main meeting programme.

• An accompanying exhibition in the purposebuilt hall alongside posters, food and drink; • A community workshop space at the heart of the exhibition, with many groups hosting meetings and running workshops.

• An event dinner with networking at the Rum Warehouse.

https://www.elmi2024.org/

37

Journal of

Microscopy The Journal of Microscopy publishes top quality research articles, review articles and Hot Topic papers covering all aspects of microscopy and analysis.

This includes cutting-edge technology and innovative applications in physics, chemistry, material and biological sciences.

You can read the latest Early View papers online at www.journalofmicroscopy.org They include:

Multimodal optical mesoscopy reveals the quantity and spatial distribution of Gram-positive biofilms in ex vivo tonsils

Megan Clapperton,Tash Kunanandam, Catalina D. Florea, Catriona M. Douglas, Gail McConnell Biofilms, which are communities of bacteria known to be more resistant to antibiotics than normal (planktonic) bacteria, are thought to play a role in diseases of the tonsil. Biofilms are difficult to study in patient tissue due to limits in standard microscopes, which compromise either on the tissue volume that can be studied or the amount of detail in the image.

We have used the Mesolens, a novel microscope lens, to visualise bacteria and biofilms in unusually large volumes of freshly excised tonsils from a local children’s hospital following tonsillectomy. Using this method, we have shown that biofilms were present in all diseased tonsils on both the surface and the interior of the tonsil.This new way of studying diseases in human tissue may prove useful in understanding the role of biofilms in other diseases and infections.

ORIGINAL ARTICLE (Open Access)

An automated slide scanning system for membrane filter imaging in diagnosis of urogenital schistosomiasis Prosper Oyibo,Tope Agbana, Lisette van Lieshout,Wellington Oyibo, Jan-Carel Diehl, Gleb Vdovine

38

ISSUE 73 MARCH 2024

Traditionally, automated slide scanning involves capturing a rectangular grid of field-of-view (FoV) images which can be stitched together to create whole slide images, while the autofocusing algorithm captures a focal stack of images to determine the best in-focus image. However, these methods can be time-consuming due to the need for X-, Yand Z-axis movements of the digital microscope while capturing multiple FoV images. In this paper, we propose a solution to minimise these redundancies by presenting an optimal procedure for automated slide scanning of circular membrane filters on a glass slide. We achieve this by following an optimal path in the sample plane, ensuring that only FoVs overlapping the filter membrane are captured. To capture the best in-focus FoV image, we utilise a hillclimbing approach that tracks the peak of the mean of Gaussian gradient of the captured FoVs images along the Z-axis. We implemented this procedure to optimise the efficiency of the Schistoscope, an automated digital microscope developed to diagnose urogenital schistosomiasis by imaging Schistosoma haematobium eggs on 13 or 25 mm membrane filters.

Our improved method reduces the automated slide scanning time by 63.18% and 72.52% for the respective filter sizes. This advancement greatly supports the practicality of the Schistoscope in large-scale schistosomiasis monitoring and evaluation programs in endemic regions.This will save time, resources and also accelerate generation of data that is critical in achieving the targets for schistosomiasis elimination.

the mesophyll cross sections in a scanning electron microscope.We applied a reduction of watering from 60 to 40 to 20 mL per day and investigated two time points, 7 and 12 days, respectively. Interestingly, the overall thickness of leaves increased in water stress conditions. Our results showed that the palisade and spongy layers behaved differently under varying watering regimes. Moreover, the results showed that this method can be used to image leaf sections after drought stress without the risk of artefacts or swelling caused by contact to liquids as during chemical fixation.

ORIGINAL ARTICLE (Open Access)

THEMED ISSUE ARTICLE (Open Access)

A rapid freezing method to determine tissue layer thickness in drought-stressed leaves Maryam Alsadat Zekri, Carina Leimhofer, Nicole Drexler, Ingeborg Lang Plants have been affected by water stress ever since they settled on dry land. In severe and persisting drought, plant leaves are wilting. However, a documentation at the anatomical level of the minute changes that occur before wilting is challenging.

On the other hand, understanding the anatomical alteration in plant leaves with respect to water stress provides a stronger basis to study molecular and submolecular processes through which plants enhance drought tolerance. In this work, we applied an affordable method to visualise mesophyll layers of Arabidopsis thaliana cell lines without preparation steps that would alter the volume of the cells. We rapidly plunge-froze the leaves in liquid nitrogen, cut them while in the N2 bath, and immediately imaged

The use of fluorescence lifetime imaging (FLIM) for in situ microbial detection in complex mineral substrates Yekaterina Chmykh, Jay L. Nadeau The standard method of bacterial enumeration is to label the cells with a fluorescent dye and count them under high-power fluorescence microscopy.

However, this can be difficult when the cells are embedded in soil and rock due to fluorescence from the surrounding minerals and dye binding to ambiguous features of the substrate. The use of fluorescence lifetime imaging (FLIM) can disambiguate these signals and allow for improved detection of bacteria in environmental samples.

ORIGINAL ARTICLE A multiangle polarised imaging-based method for thin section segmentation Yan Chen,Yu Yi,Yongfang Dai, Xiangchao Shi The most crucial task of petroleum geology is to explore oil and gas reservoirs in the deep underground.

As one of the analysis techniques in petroleum geological research, rock thin section identification

39

accurately fit the boundaries of mineral particles in sandstone images than existing image segmentation methods. Additionally, when applied in actual production scenarios, our method exhibits excellent performance, greatly improving thin section identification efficiency and significantly assisting experts in identification.

ORIGINAL ARTICLE

Atomically resolved a novel, compact and stiff scanning tunnelling microscopy in cryogen-free superconducting magnet Behnam Esmaeilzadeh, Muhammad Touqeer, Liu Junwei, Shaofeng Zheng,Tao Geng,Yubin Hou, Qingyou Lu

method includes particle segmentation, which is one of the key steps. A conventional sandstone thin section image typically contains hundreds of mineral particles with blurred boundaries and complex microstructures inside the particles. Moreover, the complex lithology and low porosity of tight sandstone make traditional image segmentation methods unsuitable for solving the complex thin section segmentation problems.

This paper combines petrology and image processing technologies. First, polarised sequence images are aligned, and then the images are transformed to the HSV colour space to extract pores. Second, particles are extracted according to their extinction characteristics. Last, a concavity and corner detection matching method is used to process the extracted particles, thereby completing the segmentation of sandstone thin section images. The experimental results show that our proposed method can more

40

ISSUE 73 MARCH 2024

We present the design and performance of a novel scanning tunnelling microscope (STM) operating in a cryogen-free superconducting magnet.

Our home-built STM head is compact (51.5 mm long and 20 mm in diameter) and has a single arm that provides complete openness in the scanning area between the tip and sample. The STM head consists of two piezoelectric tubes (PTs), a piezoelectric scanning tube (PST) mounted on a well-polished zirconia shaft, and a large PT housed in a sapphire tube called the motor tube. The main body of the STM head is made of tantalum. In this design, we fixed the sapphire tube to the frame with screws so that the tube’s position can be changed quickly.To analyse the stiffness of the STM head unit, we identified the lowest eigenfrequencies with 3 and 4 kHz in the bending modes, 8 kHz in a torsional mode, and 9 kHz in a longitudinal mode by finite element analysis, and also measured the low drift rates in the X–Y plane and in the Z direction. The high performance of the home-built STM was demonstrated by images of the hexagonal graphite lattice at 300 K and in a sweeping magnetic field from 0 T to 9 T. Our results confirm the high stability, vibration resistance, insensitivity

to high magnetic fields and the application potential of our newly developed STM for the investigation of low-frequency systems with high static support stiffness in physics, chemistry, material and biological sciences.

show that both microscopes offer slightly different advantages, and the choice between them depends on the specific requirements and goals of the user.

Submit to the Journal of Microscopy

TECHNICAL NOTE

Comparison of holotomographic microscopy and coherencecontrolled holographic microscopy Vera Chvalova,Tomas Vomastek,Tomas Grousl

1.

No submissions fees

Quantitative phase imaging (QPI) is a powerful tool for label-free visualisation of living cells. Here, we compare two QPI microscopes – the Telight Q-Phase microscope and the Nanolive 3D Cell Explorerfluo microscope. Both systems provide unbiased information about cell morphology, such as individual cell dry mass, perimeter and area. The Q-Phase microscope uses artefact-free, coherence-controlled holographic imaging technology to visualise cells in real time with minimal phototoxicity. The 3D Cell Explorer-fluo employs laser-based holotomography to reconstruct 3D images of living cells, visualising their internal structures and dynamics. Here, we analysed the strengths and limitations of both microscopes when examining two morphologically distinct cell lines – the cuboidal epithelial MDCK cells which form multicellular clusters and solitary growing Rat2 fibroblasts. We focus mainly on the ability of the devices to generate images suitable for single-cell segmentation by the built-in software, and we discuss the segmentation results and quantitative data generated from the segmented images. We

2.

No page or colour charges

3.

No page limit

4.

Simple online submission

5.

Helpful, friendly editorial team

6.

Average time from submission to first decision is less than 50 days

7.

High readership figures

8.

Online tracking system – authors can easily check the status of an article in production and receive emails at key stages

9.

Rapid publication with Early View papers published online in advance of print, significantly shortening time from acceptance to publication

10. Free electronic offprints

Journal of Microscopy App Available for iPhone and Android

Search for Journal of Microscopy on the App Store or Google play and access your personal or institutional subscription wherever you are, whenever you want.

Submit online at https://mc.manuscriptcentral.com/jmi View the Guidelines for Authors and full submission details online at:

www.journalofmicroscopy.org 41

R E P O RT

The 18th International Conference IEEE Nanotechnology Materials and Devices Conference (NMDC 2023) Paestum, Salerno, Italy 22 – 25 October

T

he 18th International Conference IEEE Nanotechnology Materials and Devices Conference (NMDC 2023) was held in the pleasant environment of the ancient city of Paestum, Salerno, Italy, on October 22-25, 2023.

Paestum is a place of exceptional artistic and natural beauty, with wonderful sea beaches as pristine as a bioreserve, nearly untouched by pollution caused by the present day public transportation system. It is known especially for the ruins of Paestum, a UNESCO heritage site famous for its three ancient Greek temples in the Doric order dating from about 550 to 450 BC. NMDC is a flagship conference series of the IEEE Nanotechnology Council (NTC), focusing on research advances in the fields of nanoscience and nanotechnology. The conference offered a great opportunity for researchers working on various aspects of nano-materials and devices, worldwide, to showcase their research on such a big platform. An important pre-conference event – ‘2023 IEEE NTC Forum on Nanomechanics and Machine Learning’ on the utilization of Machine Learning in science and technology, very relevant to today’s research, was also held on 21-22 October 2023 at the same venue.

As for the NMDC statistics, the conference

full papers were accepted for publication in IEEE

witnessed 351 attendees from 39 countries. There

Xplore with a special issue on IEEE NMDC 2023 in

were eight plenary, 48 keynote, 146 invited, 113

the journal of IEEE Transactions on Nanotechnology

contributed talks and 36 poster presentations

(TNANO).

in six parallel sessions. There were four special sessions on Nanoporous materials, Neuromorphic & unconventional computing, 2D and 3D printed electronics and Nano-enabled Icephobicity. The conference started off with an excellent plenary talk by professor X. Jiang from NC State University on Micro/Nanotechnology Enabled Intravascular Ultrasound Imaging and Therapy. A total of 113

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ISSUE 73 MARCH 2024

The most interesting sessions for me were the use of machine learning in the fabrication and characterisation of materials and on the structure-property relationship derivations from advanced electron microscopy (TEM and SEM) of materials. It was quite fascinating to know that through machine-learning-based techniques the evaluation of the complex physical properties of

these, there was always something interesting to learn in the ongoing parallel sessions throughout the conference.

I am grateful to the conference committee for selecting my paper “Direct Synthesis of Silicon Wires on a polymer substrate without the substrate heating using Hot-Wire-Enabled VLS method” for oral presentation. One of widely used bottomup methods of synthesis of Silicon wires (SiWs), the Vapor-liquid-solid (VLS) method, essentially poses limitations for their direct integration on low operating temperature (~150 °C) polymeric substrates such as Polyethylene Naphthalate (PEN) substrate. In the presented work we were able to show that direct integration of Si wires is feasible even without substrate heating via Hot-WireEnabled VLS method, a variation of conventional VLS method, in which the heated filament (the Hot-wire) plays a critical role in enabling the synthesis of the wires at sufficiently low substrate temperatures. In this way, direct integration of SiWs can be done virtually on any substrate. This work opens up avenues for replicating existing SiWsbased devices on low-melting-point polymeric substrates in a facile and cost-effective manner. My Nitin Arya at the conference centre.

nanostructured materials could be done with only a fractional computational cost of conventional DFTbased solutions, cutting down the time taken from months to tens of hours. In a talk by Dr. R. Arenal

work was well received

and appreciated by many

researchers. A full paper of the presented work can soon be referred in IEEE Xplore.

On the social front, there were four events including three conference dinners and a tour to

it was exciting to know about improved capabilities of TEM combining the imaging and spectroscopy for studying the atomic structure and the optoelectronic properties of different inorganic 1D and 2D nanostructures. In his talk, he presented his work on an in-situ thermal reduction of Graphene Oxide. On the other hand, M. Zaghloul discussed an implementation of dynamical imaging of MEMS by time-resolved scanning electron microscopy in his talk. He showed that obtaining stroboscopic movies of MEMS resonator by sequential recording of secondary electron signals is possible. Apart from

The Second Temple of Hera within the ancient Greek city of Paestum, a UNESCO World Heritage site.

43

Nitin delivers his talk.

the Archaeological Park of Paestum. These events

on the use of machine learning in nanotechnology.

provided

excellent opportunities to interact

The tour to the archaeological site quenched my

informally with researchers from across the

long-standing thirst to know about ancient Greek

world one of the best ways of networking. One

and Roman cultures. The conference was successful

event worth mentioning here is ‘Young Professional

in serving its purpose of providing a platform for

Social Event’ in which a few senior researchers were

international exposure to scholars’ research work,

also present to guide doctoral students on career prospects in research in Europe. I benefitted immensely from this event.

I was overwhelmed by the great atmosphere of cordiality and hospitality showed by the conference organisers. IEEE NMDC 2023 covered a broad spectrum of topics related to nanoscience and technology, from ‘Application of nanotechnology in

medical

sciences

(nano-biomedicine)’

to

‘Nanotechnology ethics’ with an extra 2-day forum

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ISSUE 73 MARCH 2024

opportunities for networking and information about career prospects in research in Europe.

I would like to thank the Royal Microscopical Society and Indian Institute of Technology Bombay India for the travel grant, without which it would have not been possible for me to attend this conference.

Nitin Arya Indian Institute of Technology Bombay, India

New Member Welcome The Royal Microscopical Society would like to welcome our new members who have joined us in the last three months. We hope they enjoy a long and rewarding membership with the RMS. Mr Jagen Burke Dr Bev Goward Dr Kevin Conway Lisa Pang John Maddock Mx Arman Yousefi

Dr Michael Macey Mr Warren Hatch Dr Mark Willett Mr Stuart King Dr Wanrong Geng Alan Marron

Ms Rebecca Betts Miss Katie Sharrocks Mr Ishfaq Mir Dr Liene Spruzeniece Dr Konstantinos Kalyviotis Dr Samuel Johnston

If you know of anyone who might be interested in becoming a member of the Royal Microscopical Society and if you would like us to contact them, please send their details to our Membership Administrator, Debbie Hunt – debbie@rms.org.uk Application forms are available to download at www.rms.org.uk/membership Don't forget you can now log into the RMS website and check your membership status, renew and download receipts. If you have never logged into the RMS website, please enter the email address that is linked to your membership and then click 'forgotten password'.

If you have any queries or questions about your membership please contact Debbie Hunt debbie@rms.org.uk

‘Go with the flow’, by Anne Weston,The Francis Crick Institute. A shortlisted entry to our 2023 Scientific Imaging Competition, this is an SEM image of newt larvae ciliated cells. Amphibian larvae respiration mostly takes place through skin so cilia driven fluid flow may be critical to continually replenish oxygenated water. Researchers are developing amphibian ciliated skin cells as a model system to understand development and ability of ciliated cells to generate directed fluid flow.

45

NEWS Dr Peter O'Toole.

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ISSUE 73 MARCH 2024

From the RMS President Dear Readers, It feels like time has really flown since our December issue of infocus - despite (or perhaps the result of) so much having taken place.

During January and February, we had three highly successful events in the form of UK Light Microscopy Facilities Meeting, Flow Facilities Meeting, and Electron Microscopy UK & Ireland (EM-UKI). All these meetings were geared towards helping microscopists in their day jobs, and certainly made for a highimpact start to the year, with each attracting around 100 to over 180 attendees. I was fortunate to be able to attend all three of these, and the community spirit and networking was second-to-none. That’s a huge credit to both the local organisers and the attendees themselves.

Our new Chief Executive Sali Davies has been with us now for several months, and it is great to see the impact she is making. There is a real sense of energy and excitement around our activities, and I’m really looking forward to our next round of committee meetings. I think we will see the Society evolving at pace over the next couple of years, and as always, we would love to hear from any members who have any ideas or thoughts about how the RMS can be active in supporting microscopy. It’s important to emphasise that this is your Society, and – as has been the case since 1839 – its work is entirely dependent on the input from its membership.

Our biggest event on this year's horizon is, of course, elmi2024, taking place in Liverpool from 4 – 7 June.

We are very honoured to have been tasked with delivering this year’s instalment of the meeting series, which has been staged at locations across Europe since 2001. With a likely attendance of more than 400, and a buzzing atmosphere, this promises to be a truly special occasion. The venue - ACC Liverpool - is a purpose-built convention centre in the heart of the city on the King's Dock. It looks fantastic, and I can’t wait to see all the preparations coming to fruition in early June.

We also have a wide range of other events lined up throughout 2024, and I would urge readers to visit our event calendar to see what’s on offer; we hope there is something for everyone.The RMS is a very inclusive society, and our activities cover all aspects and branches of microscopy and cytometry – plus many closely associated techniques and disciplines such as mass spectroscopy and X-ray imaging.The RMS is always keen to bring new technologies under its umbrella and to extend its support for the communities using them.

I look forward to seeing you at elmi2024 – or perhaps one of our other events later this year.

Best wishes, Dr Peter O’Toole, RMS President

47

NEWS

Professor David Stephens receives RMS Scientific Achievement Award We are delighted to announce Professor David Stephens as the latest recipient of the RMS Scientific Achievement Award.

David has been a group leader in the School of Biochemistry at University of Bristol since 2001. He is an outstanding cell biologist who uses advanced light microscopy and electron microscopy to study the secretory pathway. His work spans over

30 years of excellence in research with over 100 publications.

In 2005 he made the key finding that membranes of the early secretory pathway can be linked to microtubules and motor proteins for subsequent organisation and movement to the Golgi apparatus. He has also made many contributions to the understanding of cilia biology, including work identifying the subunit composition and role of the dynein-2 motor protein complex in the formation and maintenance of primary cilia.

In 2018 his work, David provided an important insight into the process of procollagen trafficking, revealing a short-loop pathway from the ER to the Golgi, without the use of large carriers. Further work has defined roles for early secretory pathway proteins, including giantin and TANGO1, in the processing and secretion of procollagen. David continues to publish in these areas with some exciting new publications coming up in 2024.

As well as driving his own successful research group, David has been a collaborator on many research projects, generously giving of his time and mentorship to support the cell biology community.

During his research career he has held funding from UKRI and The Wellcome Trust, with a significant amount dedicated to the provision of advanced microscopy platforms. Microscopy has always underpinned David’s research and he was an enthusiastic member of the Royal Microscopical Society Life Sciences Section for several years, supporting meetings including the Microscience Microscopy Congress (mmc).

David Stephens, with his RMS Scientific Achievement Award.

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ISSUE 73 MARCH 2024

David has been an active member of the British Society for Cell Biology and the British Society for Matrix Biology and is a Fellow of the Royal Society of Biology. He has also served as a highly regarded editor with the Journal of Cell Science since 2015.

David has also invested considerable time as a grant panel member in the UK and internationally, as well as serving on the Council of UKRI-BBSRC.

Journal of Microscopy announces Early Career Researcher ‘Best Paper’ prize winners Congratulations to Dr Charlotte Pain and Dr Alex Robinson!

The Editors of the Journal of Microscopy are pleased to announce the winners of the inaugural ‘Best Paper’ awards for Early Career Researchers in 2023.

Each prize was £200 and was judged by General Editor Professor Michelle Peckham, Deputy Editor Professor Pete Nellist and our team of Scientific Editors.

For life sciences, Dr Charlotte Pain has been awarded ‘Best Paper’ for the paper intER-ACTINg: The structure and dynamics of ER and actin are interlinked. Charlotte is a post-doctoral researcher in the department of Biological and Medical Sciences at Oxford Brookes University.

For physical sciences, Dr Alex W. Robinson has been awarded ‘Best Paper’ for the paper Towards real-time STEM simulations through targeted subsampling strategies. Alex is a post-doctoral research associate at the University of Liverpool, and Research Lead at SenseAI innovations.

Life Sciences winner Scientific Editor Dr Ulla Neumann said: “The study by Charlotte Pain and colleagues beautifully shows the intricate relationship between the actin cytoskeleton and the endoplasmic reticulum in Pain.

Charlotte plants.

Highquality confocal microscopy combined with solid image analysis and data quantification clearly show that changing one of the two partners in the plant ER-actin relationship inevitably alters the other. It also makes the point that the choice of a specific actin marker is crucial when it comes to studying organelle structure and dynamics. Congratulations to Charlotte and her

colleagues for winning the ERC Best Paper prize in the Life Sciences.”

Professor Peckham added: “This is comprehensive study that demonstrates the strong interaction between the actin cytoskeleton and the endoplasmic reticulum (ER) in plants. The images are great, have been expertly quantified, and the work additionally demonstrates how choice of fluorescent probe for actin and ER in important in avoiding off target effects.”

Physical sciences winner Scientific Editor Professor Mark Rainforth said: “Scanning transmission electron microscopy images can be complex to interpret at the atomic scale as the contrast is sensitive to many variables such as sample thickness, composition, the presence of defects, and of course, aberrations. This paper took a new approach where the manner in which the signal was sampled was changed, Alex Robinson. providing a more accurate and efficient method for STEM simulations. The paper won because of the novelty and quality of the work undertaken.”

Professor Nellist added:“This paper brings together simulations of scanning transmission electron microscope (STEM) images with the emerging field of compressive sensing. Simulations of images are a vital tool to allow the inversion of STEM data to meaningful, quantitative measurements.

These simulations are, however, computationally demanding which can limit the range of models and parameters that can be explored. Compressive sensing has been used for in-painting data from sub-sampled experimental data, but the innovation in this paper is its use for sub-sampling of modelling parameters in simulations with the aim of making simulations real time.” www.journalofmicroscopy.org.uk

49

NEWS

RMS Summer Studentships 2024 - still time to apply! £2,000 awards available for students carrying out summer projects

The deadline for applications is 31 March 2024.

Applications for the RMS’s fantastic Summer

by the student by the end of the period of study

Studentship scheme are now being invited.

and submitted to the RMS. Students will also be

Up to six studentships of £2,000 are offered

asked to do a two-to-five-minute recording briefly

to

talking about their experience for the RMS YouTube

undergraduates

every

year, split

evenly

The Studentship is offered on the understanding that a 500-word project report is completed

between physical sciences, biological sciences and

Channel.

interdisciplinary projects.

Students who are in their first or final year are not

Applications for our Summer Studentships must

eligible. The person making the application must

include a significant microscopy component and

be a Member or Fellow of the Royal Microscopical

should be submitted by a suitable host academic on

Society.

behalf of a student.

Find out more and apply

Ben Watson (University of Strathclyde) completed an RMS Summer Studentship in 2022. You can read his report – and others – in our Article Backfile.

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ISSUE 73 MARCH 2024

Journal of Microscopy: Call for Papers for new Special Issue Microscopy and Infectious Diseases (host-pathogen interaction and pathogens structure) The Journal of Microscopy is pleased to announce a

Read our guidelines for authors

new special issue featuring papers on “Microscopy

Deadline: September 30th, 2024

and Infectious Diseases (host-pathogen interaction and pathogens structure)”.

The special issue will be guest edited by Dr Leandro

If you have any questions, please contact Editorial Office Manager Jill Hobbs, journaladmin@rms.org. uk

Lemgruber, Head of the Cellular Analysis Facility, University

of

and

Mariana

De

Niz, The

Glasgow, UK,

Feinberg School of

Medicine,

Nor thwestern University, USA. We welcome submissions for this issue and papers can be Reviews, Methods and Protocols or Primary Research articles.

RMS now administering membership for Association of Clinical Electron Microscopists (ACEM) Contact us to renew your ACEM membership

membership applications - following confirmation of membership by the ACEM committee. New and

The RMS is delighted to be providing membership

‘lapsed’ members can apply for membership via the

administration support for the Association of

website at www.acem.org.uk/membership.

Clinical Electron Microscopists (ACEM).

The ACEM membership fee is now £35 including

The Society has taken on responsibility for

a nominal RMS administration fee. To renew your

maintaining membership records, receiving funds and issuing renewals. It will also process new

ACEM membership, please contact Debbie Hunt.

51

NEWS

RMS supporting Women in STEMM Summit 2024 Chief Executive Sali Davis joins line-up of speakers at ITSHER Scientific Conference (7-8 March, London and Dublin)

Anna Baldycheva.

The RMS is proud to support the upcoming ITSHER 2024 conference, part of the Women in STEMM Summit 2024.

be the opportunity to watch the sessions and panel discussions interchangeably through live stream. The event has been organised by Professor Anna Baldycheva (Exeter University) who is also Chair of the RMS Engineering, Physical & Material Sciences Committee.

The scientific programme is carefully curated to offer a blend of powerful talks and enlightening poster presentations, showcasing the incredible research and innovations spearheaded by female Sali Davis.

scientists and engineers across the UK and Ireland.

Topics include all STEMM disciplines: Physics,

The groundbreaking event will bring universities

Engineering,

and businesses in the UK and Ireland together

Health Sciences, Life Sciences, and Material Science.

to celebrate science, research and innovations by

Moreover, the programme is enriched by special

women across all disciplines in STEMM.

sessions and panel discussions devoted to career

RMS Chief Executive Sali Davis will join an exciting cast of speakers at the dynamic, interdisciplinary conference, taking place simultaneously at the Institute of Physics (IOP), London, and Trinity College Dublin (TCD), Dublin. There will also

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ISSUE 73 MARCH 2024

Mathematics,

Computer

Science,

development in academia and industry for female professionals, as well as Innovation Sessions with the opportunity to receive feedback from industry practitioners.

Find out more and register

RMS team goes behind the scenes at London’s Natural History Museum The RMS would like to give special thanks to Dr Alex Ball and Dr Alex Sossick for hosting a special staff visit to the Natural History Museum in London on 1 February.

The RMS team pose for a group photo on the steps inside the famous Hintze Hall.

The team was given a ‘behind-the-scenes’ tour to take a look at some of the fascinating science and microscopy taking place at the iconic venue – where both ‘Alexes’ are currently based.

Dr Alex Ball holds a model of the famous 'Dippy' dinosaur skull, while giving a talk to RMS staff.

From scanning microscopy and 3-D modelling, to conservation and fossil preparation, staff were given a taste of the wide-ranging research currently being undertaken.

There was even time to view some of the museum’s extensive hoard of animal specimens stored in alcohol in the Museum’s spirit collection. A visit to the Wildlife Photographer of the Year exhibition capped off a memorable away-day.

Once again, our thanks to Alex and Alex, for taking time out from their busy schedule to make this Staff were given a chance to explore some of the electron microscopy taking place at the museum.

possible.

53

IMPRO SUCCESS TRAINING MOTIVAT WORK

OVE The RMS Mentoring and Application Scheme: Nurturing Microscopists across the world Joëlle Goulding and Georgina Fletcher, RMS Mentoring Working Group

E

The RMS mentoring scheme is unique as it is crafted by microscopists for microscopists. Indeed, this is the reason why it came about in the first place, as we aimed to address the distinctive needs of individuals in this specialised field. Originating from conversations among Joelle, Scott, Georgina, Paul, and Alex, aka “the Mentoring Working Group”, we recognised the lack of microscopist-focused guidance. Given the niche nature of our roles, seeking career or specialist advice often means looking beyond our immediate workplace or even country, and building such networks can be challenging.

So, in 2022 and 2023 we conducted a pilot of the mentoring scheme. This provisional programme encompassed 1) personal mentoring for soft skills and 2) application mentoring pairing microscopists with specific instrument or software expertise. Drawing on their professional connections, the Mentoring Working Group matched together eight pairs, catering to early career as well as more established microscopists that were looking for a range of diverse goals, from transitioning to industry to developing core facility management skills. The feedback was very positive, with some examples overleaf…

55

“It’s been great so far. I am very lucky to have a mentor that is passionate about helping me.

She is a brilliant listener and gives me advice that I don’t get from academic supervisors.

The insight into industry has been super.”

“Just thanks! It’s been very good so far!”

“Early days, but I’m optimistic about the insights and interest the mentor has shown” Buoyed by the pilot’s success, we officially launched

and successful pairings, including several on the

the scheme in July 2023, with plans for biannual

application coaching stream. Applicants hail from

pairing rounds. Applicants are asked to provide

various corners of the globe, expanding our

detailed information about their mentoring needs,

network of both mentees and mentors.

job role, and career stage, with location details being non-restrictive, as most interactions occur virtually. The working group diligently reviews applications, aiming to find the ideal mentor either from our growing database or the wider microscopy

Below, some of our pilot cohort reflect on their experiences of mentoring, what they have learned and advice for anyone thinking of getting involved themselves:

community.

The mentorship experience is rewarding for both parties, fostering confidence, career growth, and transferable skills like leadership, empathy, networking, and communication. We encourage more mentors to join our database by completing the online application form. Once pairs are matched, we introduce them via email along with some mentoring resources kindly provided by the Francis Crick Institute. While the pairings are initially designed for six-to-12 months, there’s no strict limit, allowing connections to flourish as long as both parties desire. Periodic check-ins and feedback questionnaires contribute to continuous improvement.

Our first cohort, initiated from the July-August 2023 call, is now 4-6 months into their pairing.

The response to the inaugural call surpassed our

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expectations, with

ISSUE 73 MARCH 2024

numerous

applicants

Charlotte Clews: Mentee (Application mentoring) Roslin Institute, University of Edinburgh

Why did you decide to get involved?

to be part of.

As a scientist with a fascination for microscopy and

Is there any advice you would give for

the recent technical and methodical developments

anyone thinking of becoming a mentor /

in bio-imaging, it seemed a perfect fit for me to get

mentee?

involved with a fantastic society like the RMS, and also be able to receive advice and help from experts in this field, to help me develop in my research career.

Can you describe what you have been doing and your experience so far?

Mentees: Ask all the questions you feel you need – a great part of this partnership is just getting a more well-rounded understanding of microscopy.

Mentors: Mentees often have gaps in their knowledge that even they are sometimes not aware of. Sometimes we need to go back to the

So far I have been having 1:1 meetings with my

basics to build up a better understanding of the

mentor to gain general advice about improving

techniques we’re learning/attempting. Clarity and

my skills in microscopy, and also guidance while

sometimes step-by-step methods (if relevant) are

attempting advanced imaging techniques such as

really valuable to us!

Correlative Light and Electron Microscopy (CLEM).

How do you feel you have benefitted from taking part?

Being able to ask questions (both complex and simple!) to gain a broader understanding of the field, the different techniques and different instruments available to me, has been very useful.Also just having a second opinion on my images/figures/experimental design has been very useful indeed, and helped me develop better approaches to answer my biological question.

What have you learned that will be most useful to you going forward?

Imaging and the preparation steps are often (but not always) a multi-step process. Always have extra samples, extra images, extra everything (!) to make sure that by the end you have useful, representative and reproducible data to take forward to analysis.

Why do you think the RMS scheme is important for the microscopy community?

I think microscopy can seem exciting but very daunting to get into – so having experts and a

Virginia Silio: Mentee (Personal mentoring) Confocal Technician, University College London Why did you decide to get involved?

welcoming community like the RMS ready and

I felt that my career wasn’t progressing at all,

available to help support those wanting mentoring

and I didn’t know what to do about it. I tried the

is a very valuable resource, and one I’m very happy

mentoring programme inside my institution, but

57

being in a microscope core facility is a very niche

we couldn’t connect, and I felt she didn’t understand

set of skills that is difficult to understand for an

what I did.

outsider. I saw the mentoring program in the RMS

Is there any advice you would give to

and I thought it would be a great opportunity to have input from someone senior in the field.

Can you describe what you have been doing and your experience so far? We have met using Teams several times. I am hoping we can catch up in person soon. It was very informal, we discussed my progression and what I want to achieve in the future, and my mentor gave me ideas to work with.

anyone thinking of becoming a mentor/ mentee?

My experience is only as a mentee, and I will recommend anybody thinking about going for it.

I would recommend to anybody more senior to become a mentor, as the community needs as much input as possible.

How do you feel you have benefitted from taking part?

I have learnt a lot from my mentor. I discovered some paths I didn’t think about or even didn’t know existed. During that time, I applied and changed jobs.

I was part of the Herschel program too.

I have also benefitted from having an external point of view, the input is very useful.

I also saw photos of puppies, that was a bonus.

What have you learned that will be most useful to you going forward?

We talked about different opportunities for learning new skills that will be useful for career progression (e.g., apprenticeship). Also, we discussed about group dynamics, how to navigate those dynamics, etc. He is in different working groups, so it was very useful to talk to him about that.

Peter O’Toole – Mentor

Why do you think the RMS scheme

RMS President, Head of Imaging and

is important for the microscopy

Cytometry Labs, University of York

community?

It allows contacting and connecting with people with a career more related to yours. There are other mentoring opportunities schemes out there (in the workplace, other professional bodies), but I found them very general, and our professional profile is quite different.

58

Why did you decide to get involved?

The simple answer is that I was asked to help! – but mentoring was something I had already been doing with a few people informally, as they took on new roles. I had previously mentored people I had known from conferences who contacted me afterwards for advice. I have also mentored PhD

As I mentioned, I tried the one offered at my

and post-doc students who kept in touch after

previous institution. The mentor was very nice, but

continuing their careers.

ISSUE 73 MARCH 2024

Can you describe what you have been

does take time to grow into the role, and there is

doing and your experience so far?

obviously the time commitment in general. But it

Having joined the programme as a mentor, I suddenly had multiple requests for mentorship overnight ! Sometimes I’m giving help and technical

can be very rewarding, and the self-reflection that comes from taking part is really important. From a moral perspective, I also think it is a good thing to

advice as a scientist - on what works best from a

give something back, and to help your community

science perspective; at other times it’s about the

in general.

intricacies of running a facility – from charging and HR procurement to communication with users. At that level, it is not just about microscopy, and it can involve people from outside the microscopy world.

Paul Verkade: Mentor Professor of Bioimaging, University of

How do you feel you have benefitted

Bristol

from taking part?

Why did you decide to get involved?

The meetings give the mentees a lot of enthusiasm,

Before we even discussed setting up the scheme, I

and when you hear that they feel really motivated and itching to get back and put some of the ideas you have been talking about into practice, that is very rewarding.

had been contacted by Scott Dillon who was setting up a new facility and asked if I wanted to mentor him in that process - dealing with users, career progression and those kind of issues. That was a

There’s an element of worry in case you give the

very useful experience for both of us and it was

wrong advice to someone – but overall it is a really

then only natural to be engaged when it grew into

positive process which can help both the mentor

the RMS scheme.

and mentee to come up with new ways of working.

What have you learned that will be most useful to you going forward?

Every time you mentor someone, you inevitably reflect on what you are doing yourself. Mentees will ask difficult questions and challenge your thinking, and you question yourself as to whether you are still enacting best practices. Working practices are always evolving, so that process of self-reflection is a very healthy and helpful thing.

Why do you think the RMS scheme is important for the microscopy community?

I think that sometimes people don’t have the confidence to ask for help - or to know who to ask.

That is why having a formal mentoring programme is so important – because it really empowers people to come forward and removes that barrier.

For anyone thinking of becoming a mentor, it

Can you describe what you have been doing and your experience so far?

We meet up every now and then, mainly depending on the need, and we have a chat about questions the mentee has and I try to share my thoughts and experiences from my career. It depends what kind of mentorship it is, “soft skills” or more technical.

With the soft skills you always have to remember

59

that these are my own experiences and don’t always

Why do you think the RMS scheme

translate one-to-one to another situation.

is important for the microscopy

How do you feel you have benefitted from taking part?

For me, personally, it has been a very useful exercise in reflecting on my own career - the decisions I have made, how I have learned from my experiences, why am I in the position I am now (that sounds quite grand!).

What have you learned that will be most useful to you going forward?

That the microscopy community is a very friendly and open community and because of that, it is very easy to discuss and engage with other people. It is my second family.

community?

We can learn so much from each other and we should not let that know-how go to waste.

Is there any advice you would give for anyone thinking of becoming a mentor / mentee?

Everyone has been in the situation where they started something and were not sure how to tackle this. Most likely you will have thought, who can I ask? Maybe there was someone, maybe there wasn’t.

Both are the perfect reason to get involved, because there are a lot of microscopists out there very willing to share their experience.

Find out more about the RMS Mentoring and Application scheme

Submit to infocus infocus welcomes submissions of articles of general interest to microscopists.

You provide the text and images and we take care of the rest. It’s the ideal way to share your work with the microscopical community.

Full submission information and guidelines are available at www.infocus.org.uk.

To submit an idea or if you have any questions about the process please email the Editor (editor@infocus.org.uk)

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C O M PA N Y N E W S

Updates from FocalPlane 2023 was an exciting year for FocalPlane, our online microscopy community site. We’ve seen upgrades to the site, new contributors and fresh content from our regulars! The number of people using and contributing to the site is also growing, as we continue to reach out to the global community of imaging scientists, engineers, bioimage analysts and cell biologists.

We recently implemented two main upgrades to the site: a new partnership with MicroscopyDB, and a new look for the site thanks to our image competition finalists. The partnership with MicroscopyDB means that our events calendar and jobs board are now powered by MicroscopyDB, making the calendar and board essential viewing for planning your conference travel or finding for a new position. On the FocalPlane homepage, we have released a series of banner images which now appear alongside our characteristic lens apertures.

We thank Marie-Charlotte Domart & Chris Peddie, Nick Gatford, Oona Paavolainen, Rebecca Simkin and Till Stephan for sharing their images with us.

2023 was also a busy year for our FocalPlane correspondents, Mariana de Niz and Mai Rahmoon.

Mariana continued her excellent ‘Latin American Microscopists’ series, while also starting a ‘Toward Global Access’ series with Constadina Arvanitis. Mai

has been introducing us to imaging communities through interviews with their founders. We look forward to continuing to work with Mariana and Mai, and we are excited to announce our new team of correspondents, Daniel Doucet, Subhajit Dutta and Greg Redpath. Visit our website to read our announcement post introducing the three of them and find out what topics they will be writing about for FocalPlane.

We’ve also been busy planning some new initiatives for 2024 and hope to announce these soon. The easiest way to stay up to date with the latest news from FocalPlane is by getting our weekly digest or our quarterly newsletter delivered straight to your inbox. Simply register with the site and select the appropriate box.

Finally, we would like to thank all our readers, subscribers, and most of all, our contributors in 2023. Remember, FocalPlane is your site. Once registered, you are free to post, and if you have suggestions or feedback, you can contact the team at focalplane@biologists.com. https://focalplane.biologists.com

Labtech appointed as a distributor for Tomocube and Nikon Labtech International has been appointed as exclusive UK distributor of Tomocube, the innovative manufacturer of cutting-edge label-free 4D Holotomography systems. Holotomography utilises 360° quantitative refractive index measurement to build 3D holograms with high resolution and temporal specificity while minimising cellular stress.

The Tomocube range includes the HT series for live cell imaging, the HT-X1 series for higher throughput using multiwell plates, and analysis software to unleash the full potential of Holotomography.

In addition, Labtech has been appointed as one of the UK distributors for Nikon’s range of life science microscopes. Nikon is a leader in microscopebased imaging technologies for the life sciences and Labtech is delighted to be working alongside their team of UK specialists. We can supply and support your imaging needs, especially within the private sector. https://www.labtech.com

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KEYENCE VHX-970F Microscope Proves that Knowledge is Power at TDK-Lambda UK TDK-Lambda UK, a major manufacturer of power supplies to markets such as medical and industrial, is taking advantage of a newly installed KEYENCE VHX 970F high accuracy digital microscope to drive the company’s inspection and quality function. With such a reputable global brand, there is no margin for failures.

Part of Japanese multinational electronics giant TDK Corporation, TDK-Lambda UK is Britain’s largest designer and manufacturer of both standard and configurable AC-DC and DCDC power supplies. The company’s 350-employee facility in Ilfracombe is home to its manufacturing, new product development, sales and service functions. A key part of delivering business success at TDK-Lambda UK is component engineering.

“As a department, we’re responsible for component analysis; looking at alternative or new components that we test for robustness and conformity to data sheet specifications,” explains Component Engineering Manager Dan Massey. “We also look after supplier quality from our contract

manufacturers, while further responsibilities include goods inwards, sample inspection and compliance with RoHS.” TDK-Lambda UK’s previous KEYENCE microscope served the company well for around 15 years but it upgraded to keep pace with the latest innovations and improvements.

“We saw the KEYENCE VHX970F high-accuracy digital microscope and knew it would meet our requirements,” says Massey. “Also, staying with KEYENCE meant we could continue using our existing lenses. It was an easy choice.”

As an option, the company invested in a digitally controlled Z-axis for the VHX-970F that it did not have previously.

“It allows us to automate the focus, assisting ease of use,” he says. “Supported by the microscope’s impressive depth of field we can also use the digitally controlled Z-axis to take multiple images from different height perceptions if we have a particularly awkward or complex component to inspect.” www.keyence.co.uk/TDK

Easyscan 2 AFM end of service

The EasyScan 2 system with its compact design had been a very popular instrument until Nanosurf discontinued its production back in 2013. Now, 10 years later, we are facing more and more challenges servicing these instruments due to the lack of availability of spare parts. Nanosurf therefore decided to officially end the service for the Easyscan 2 AFM system.

Nevertheless, we strive to provide our customers with as much support as possible. You may still

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ISSUE 73 MARCH 2024

contact us in case of service requirements, and we will evaluate on a case-by-case basis if service of the device is still possible.

We recommend to EasyScan 2 AFM users to consider an upgrade to FlexAFM, one of the most established research AFMs available. The scan head recently received a design update that provides additional benefits: • Footprint now matches DriveAFM: the result is a shared accessories universe across systems.

• Larger number of functional accessories.

• Improved top/side-view camera.

• FlexAFM can be run with the high-end CX controller, bringing you improved performance. www.nanosurf.com

Queensgate awarded funding for fourth research project with NPL Queensgate, a brand of Prior Scientific, has won funding from Innovate UK, the UK’s innovation agency, for a fourth nanopositioning research project as part of Analysis for Innovators (A4I) program (Round 11).

This will be the fourth project Queensgate has won funding for through A4I, and once again, they will be working with the dimensional nano and sub-nano-metrology team at the National Physical Laboratory (NPL). This project will qualify the performance of Queensgate’s velocity control and special correction capabilities when applied to multi-directional motion profiles.

As Queensgate’s product manager, Craig Goodman, explained, ‘Our aim is to provide improved performance for atomic force microscopy, nanolithography and fast optical alignment systems.’

Queensgate has already developed a number of unique capabilities such as velocity control and spatial correction which allow very high-speed raster scanning with high precision. This project will help the company to refine these capabilities further.

Previous projects that Queensgate has partnered with NPL on include Spatial positioning correction for multi-axis nanopositioning stages; Improved linearisation of single-axis piezo nanopositioning stages; and developing a mathematical model for multi-axis control.

Innovate UK has released a case study video about the first, completed, project: Improving micron scale accuracy. www.prior.com www.nanopositioning.com

EM Resolutions Expands at Keele University Science and Innovation Park

We are thrilled to unveil our new, expanded headquarters at Keele University Science and Innovation Park. Our state-of-the-art facilities now include a production lab and spacious offices, allowing us to take our services to the next level.

Since relocating to Keele’s campus, our dedicated team has expanded, and our capabilities have multiplied. This means we can better serve our valued customers. But we’re not stopping there.

With larger office and laboratory spaces, EM Resolutions is now even more equipped to provide top-quality accessories to as many researchers as possible. Whether you’re a seasoned scientist or a curious newcomer, our expertise and commitment remain unwavering.

We extend our heartfelt gratitude to our loyal customers and the Science and Innovation Park.

Your support has been instrumental in our journey.

For those who haven’t yet collaborated with us, allow us to introduce ourselves: EM Resolutions is a leading UK supplier specialising in TEM Microscopy accessories. Our focus includes the manufacture and supply of support films, and we are proud to be the sole UK distributor of UltrAuFoil®, meticulously crafted by Quantifoil®.

Join us as we continue to empower scientific discovery. Together, we’re shaping the future—one support film at a time. www.emresolutions.com

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C O M PA N Y N E W S

Oxford Instruments plc acquires First Light Imaging SAS

Oxford Instruments, a leading provider of high

Richard Tyson, Chief Executive Officer, Oxford

technology products and services to industry and

Instruments plc, said: “We are delighted to welcome

scientific research communities, announces the

the talented First Light team to Oxford Instruments.

acquisition of First Light Imaging SAS (“First Light”),

Like us, they have a reputation and track record

a scientific camera specialist.

for quality, innovation and scientific expertise,

Based near Aix-en-Provence, France, and founded

in

2011,

First

Light

and their leading range of products dovetails well with our scientific camera portfolio. Joining forces

specialises in the design and

will significantly enhance

manufacture of high-speed,

our world-class camera

low-noise scientific cameras for

infrared

and

offering to our combined

visible

customer base and drive

imaging, with applications in

First Light’s next phase

astronomy and life sciences.

It

will

become

part

of growth as part of the

of

wider Group.

Oxford Instruments Andor (“Andor”), with a common global

customer

base

spanning leading academic institutions

“Our expanded portfolio will help us accelerate our customers’ roadmaps in the life science

and commercial companies. Its cameras are highly

market and in physical sciences, a long-standing area

complementary to Andor’s existing scientific

of strength for Oxford Instruments.”

camera portfolio, with the acquisition providing opportunities to enhance both companies’ offerings and to expand into adjacent markets.

David Boutolleau, co-founder of First Light, said: “Joining Oxford Instruments and Andor is a real opportunity for First Light Imaging. After 12 years

Under the terms of the acquisition, Oxford

building First Light as a successful independent

Instruments is paying an initial cash consideration

business, it became clear that to take the business

of €15.7m, on a cash-free, debt-free basis, with a

to the next stage of its development, we would need

further €3m cash consideration conditional on

industrial and commercial support to reinforce our

First Light’s trading performance over a 12-month

manufacturing, technology and routes to market.

period following completion. First Light’s estimated revenue for the 12 months to 31 December 2023 is €8.0m (£7.0m), with return on sales below the average for the Group.

“Our acquisition by Oxford Instruments Andor accelerates our growth and secures our future for our customers, partners and employees.” www.oxinst.com

If you would like your Company News to appear on these pages, please contact infocus Magazine at advertising@infocus.org.uk The announcements in this Section are compiled by the manufacturers. They in no way represent a recommendation by the Royal Microscopical Society for any particular instrument or equipment. The Royal Microscopical Society does not endorse, support, recommend or verify the information provided on these pages.

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NEW PRODUCTS

NuNano adds SPARK to the Oxford Instruments eStore AFM probe range

range of AFM probes alongside Oxford Instruments Asylum Research AFM instruments.

“Nano-electrical measurements are becoming increasingly important for materials characterisation, particularly for those materials being incorporated into a new generation of electronic devices” said James Vicary, NuNano Co-founder & Managing Director.

NuNano, the high-quality atomic force microscopy (AFM) probe manufacturer, is delighted to announce that their conductive AFM probe range, SPARK, are now available on the Oxford Instruments eStore.

The addition of SPARKs to the eStore reflects the expanding global distribution partnership with Oxford Instruments Asylum Research (Santa Barbara, US). The integration of SPARKs broadens the product offering, ensuring customers achieve reliable imaging every time when using NuNano’s

The SPARK range of conductive AFM probes are designed for electrical AFM measurements, such as Kelvin Probe Force Microscopy, Piezo-response Force Microscopy and Conductive AFM. Attention to quality and reliability, guarantees tip radius, while the symmetric tip profile ensures measurement precision across a wide range of nano-electrical applications.

Manufactured with the tightest dimensional tolerances in the market, SPARK conductive AFM probes offer minimal variation in spring constant and resonant frequency, setting the standard in probe reliability and performance. The SPARK series joins NuNano’s silicon AFM probe collection (SCOUT) in the Oxford Instruments eStore and direct from their regional offices. www.nunano.com

New LT14030 substage for SEM, FIB and UHV The new Kleindiek Nanotechnik LT14030 substage is designed for orthogonal positioning in atmosphere, SEM/FIB and UHV. The LT14030 replaces the previous model LT12830 and features greater stability and drift performance. Its primary use is in SEM and FIB-SEM to enhance the accuracy and functionality of standard microscope stages. It is an economical alternative to laser interferometer stages. The LT14030 uses two positional encoders per axis for automatic yaw error compensation and has an option for interferometers, should that extremely high degree

of precision be required, after all.

Applications include eBeam lithography, memory cell and particle counting, tensile measurement, forensic analysis (e.g.

GSR), metrology and failure analysis.

Sussex-based Labtech International offer the LT14030 and a full range of Kleindiek instruments in the UK and Ireland. https://www.labtech/em.com

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NEW PRODUCTS

Teledyne to showcase new scientific imaging and machine vision solutions at SPIE’s Photonics West exhibition Teledyne will showcase their newest products and solutions at SPIE’s Photonics West exhibition taking place January 30 – February 1st, 2024, in San Francisco, California.

Visitors to the Teledyne booth #327 can expect representation from the businesses within its Imaging group, including Teledyne DALSA, e2v, FLIR, Acton Optics, Judson, Lumenera, Photometrics, and Princeton Instruments.

New products introduced include: Teledyne e2v announces OnyxMax™, the next generation of its popular Onyx 1.3M low light CMOS image sensor. This new sensor has been designed for extremely low light conditions, down to 1 mLux. The combination of sensitivity and image resolution increases its range, allowing even small objects to be detected in harsh conditions.

This makes OnyxMax ideal for a wide range of applications including science, defense, traffic

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cameras, broadcast, surveillance, border control and astronomy.

Teledyne Photometrics releases the Retiga E9 camera that can capture exposures from tens of microseconds to tens of minutes, delivering detailed, high-resolution images without extraneous noise. With its stacked CMOS sensor technology developed by Sony, the E9’s square pixel array with 3.76 µm pixels and high dynamic range (>80 dB) combine to create a sensitive, versatile, and costeffective low noise camera ready for integration or benchtop use.

Teledyne FLIR IIS introduces the new Dragonfly® S USB3 series designed to accelerate the inception stage of imaging application development. The Dragonfly S series addresses the essential need of a modular, compact, and lightweight camera for atscale manufacturing, volume-based applications, and multi-camera systems. www.teledyne.com

Harnessing MicroXRF for Enhanced Mineral Exploration and Mining

The mining industry is constantly searching for innovative and efficient technologies to gain deeper insights into the composition and structure of geological materials. One technology that has emerged as a game-changer is Micro X-ray Fluorescence (MicroXRF) Spectrometry. This stateof-the-art method, derived from X-ray fluorescence spectroscopy, enables the analysis of geological specimens at a micro-scale level. By irradiating a specimen with a finely focused X-ray beam, the atoms within the sample emit characteristic secondary X-rays, which can be used to determine the sample’s elemental composition. With the exceptional precision of MicroXRF instruments, elemental maps of thin section and core samples can be obtained, with each pixel, down to 5µm diameter, containing its own unique elemental spectrum. This high-resolution mapping capability allows for further phase analysis, which is crucial in understanding the mineralogy of the samples.

MicroXRF is revolutionising mineral exploration, resource management, and quality control in the mining industry.

MicroXRF possesses a notable advantage in its non-destructive and non-invasive nature. This characteristic ensures the preservation of sample integrity and allows for consecutive analyses

without any harm or alteration to the specimen.

Real-time analysis and immediate results provided

by MicroXRF are of utmost importance in timesensitive operations within the mining industry.

Mineral exploration stands as a significant application of MicroXRF in the mining field. It facilitates enhanced geochemical analysis, enabling geologists to accurately identify potential mineral deposits. For instance, mineral alteration zones often encompass ore bodies, containing specific elements that may indicate the presence of valuable resources. By employing MicroXRF, explorers can detect these altered mineral phases and subsequently improve their predictive models for the presence of ore.

MicroXRF is an essential tool in resource management, particularly in the control of ore grade.

This advanced technology enables the rapid and precise determination of element concentrations in ores, including both valuable elements such as gold, copper, and nickel, as well as potentially harmful elements like arsenic, lead, or mercury. By utilising this valuable information, mining companies can effectively plan their extraction and processing operations while also mitigating potential environmental risks. www.qd-uki.co.uk

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NEW PRODUCTS

Excelitas Technologies Introduces X-Cite XYLIS II for Fluorescence Microscopy Excelitas Technologies Corp., a leading industrial

performance. XYLIS II uses the same speedDIAL,

technology manufacturer focused on delivering

light guides, adaptors and command set as the

innovative,

original XYLIS, allowing for a seamless transition

market-driven

photonic

solutions,

announces the X-Cite XYLISTM II Broad Spectrum LED Illumination System.

between the two systems.

XYLIS II features include:

A replacement for

• Whisper quiet operation

the current X-Cite XYLIS, XYLIS II offers

• Broad spectrum, UV and visible

several improvements,

• Light guide coupled, optimised for 3mm

including

significantly

• Power levels comparable to (or better

quieter

operation

than) the current X-Cite XYLIS

and increased optical output,

with

broadest

spectrum

• Control – manual (via

the

speedDIAL), TTL, USB “Excelitas is proud

available in a white light

LED

of our long-standing

for

commitment to

fluorescence

delivering X-Cite

microscopy

Lamp and LED

applications.

light sources for

The introduction

fluorescence

of XYLIS II comes as Excelitas Technologies

celebrates

the

20-year

anniversary of the X-Cite product brand, which has grown to be the industry standard in fluorescence illumination.

offering

exceptional

stability

and

superior

illumination uniformity to meet the unique requirements of our customers,” said Michelle Gal, Senior Product Manager, X-Cite at Excelitas.“XYLIS

Rivaling traditional arc lamps for brightness,

II builds upon the success of our 20-year X-Cite

XYLIS II is ideal for both compound and

heritage and addresses the feedback we’ve received

stereomicroscopes.

Excelitas’

from our valued customers. With optical output

patented, award-winning LaserLED Hybrid Drive® technology to overcome the LED green

improvements across its broad spectrum and

gap from the 540nm to 590nm region of the

the only thing customers will notice about XYLIS II

spectrum, where LED-only systems are significantly

is its great performance.”

It

incorporates

challenged.The powerful output and broad DAPI to Cy7 spectral range of XYLIS II offers researchers and microscope lab managers the benefits of LEDs without compromising on price, flexibility or

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microscopy,

ISSUE 73 MARCH 2024

sound levels 10dB quieter than the original XYLIS,

https://www.excelitas.com/product/xcite-xylis-ii-broad-spectrum-ledillumination-system

Imaris 10.1 Released – For Fast and Simple Segmentation Using AI Oxford Instruments - Andor has released Imaris 10.1, the latest version of its market-leading AI microscopy image analysis software. Imaris 10.1 integrates AI segmentation tools into its main image analysis workflows, improving

ease

providing

better

of

use and

and more

versatile segmentation for all researchers across life science applications.

In Imaris 10.1, a native, trainable AI pixel classifier is an integral part of its big-data-capable Surface segmentation model. Using the Imaris AI pixel classifier simplifies and improves the segmentation and object detection in challenging fluorescent datasets. In addition, it opens doors for 3D Scanning Electron Microscopy (SEM) segmentation and shape recognition.

Anna Paszulewicz, Software Manager for Oxford Instruments - Andor, says: “Imaris 10.1 AI pixel classifier delivers a whole new experience of interactivity and speed. Together with Imaris’ native

Crucially, the AI pixel classifier can be used by all

shape and distance measurements, it’s an amazing

researchers, regardless of their experience. Users

analysis and 3D visualisation tool for a wide range

can train and compute the results with the classifier

of research projects.”

on their local machine by painting the data with the efficient and modern smart brush tool. Training and the results preview are extremely fast and interactive, improving the user experience and saving researchers’ time.

The pixel classifier can be trained and utilised on 2D, 3D multi-channel and time-lapse datasets and used in a batch mode to analyse more samples of similar type.

Imaris’ goal is to bring researchers the most comprehensive visualisation and analysis software for 3D/4D and time-lapse microscopic images.

It’s notable that Imaris is inclusive for all available microscopy file formats and provides seamless conversion to native IMS, using Bio-Formats.

Users can download a 10-day free trial of Imaris to explore the software first-hand. https://www.oxinst.com/

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NEW PRODUCTS

Livecyte adds Neurite Outgrowth to its portfolio of label free assays for single cell behaviour Phasefocus label-free technology is breaking new ground with the development of its latest product, the Neurite Outgrowth assay. This pioneering new assay will aid researchers in both developmental studies investigating processes involved in the initial organisation of neuronal networks as well as the study of neuronal pathologies, such as degenerative diseases or neurotoxicity without the potential harmful effects of dyes and labels.

Livecyte revolutionises neurite outgrowth assays by producing high contrast, label-free Quantitative Phase Images (QPI) that can be automatically analysed to track cell bodies, and neurite characteristics (e.g. length and branching) during the development of an in vitro network. This enables researchers to quickly obtain metrics on their neurite outgrowth assays, without the requirement of costly fluorescent reagents which can impact the behaviour of their cells or by laborious manual analysis. By segmenting and tracking cells alongside a unique skeleton profile, Livecyte automatically quantifies neurite outgrowth metrics per cell and enables the tracking of cellular migration and organisation of cells during initial network formation, revealing trends that would have been overlooked by looking solely at population-based metrics. This cutting-edge technology leads to an increase in the physiological relevance of data

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ISSUE 73 MARCH 2024

collected more accurately recapitulating in vivo neuronal cell behaviour in vitro and ultimately improving drug development efficiency.

Key Advantages of Livecyte Neurite Outgrowth Assay: • Automated, accurate neurite detection labelfree: Livecyte employs advanced algorithms to automatically segment and skeletonise neuronal cells allowing streamlined network measurement.

• The Neuronal cell dashboard presents a wealth of metrics based on neurite characteristics, offering an invaluable depth of insight into the effect of mediators on neurite outgrowth.

• Livecyte’s ability to independently quantify neurite growth on a per cell basis eliminates any seeding density dependant differences and providing a more profound understanding of neuronal cell behaviour.

• Single-cell analysis of migration and morphology metrics alongside neurite outgrowth provides a more comprehensive way of understanding early neuronal cell behaviour.

• All this is achieved without the need to fluorescently label neuronal cells which alters their natural behaviour. https://www.phasefocus.com/ applications/neurite-outgrowth

Teledyne Photometrics extends its Retiga series of long-exposure cameras with the Retiga E9 Teledyne

of Teledyne

The Retiga E platform, using the Teledyne

Technologies, announces the release of the Retiga

Photometrics, a

part

Photometrics Citadel Sensor Chamber, isolates the

E9 camera that can capture exposures from tens

sensor from the environment and allows air cooling

of microseconds to tens of minutes, delivering

down to below -20° C. The platform is designed

detailed, high-resolution images without extraneous

for easy device integration into instrumentation,

noise. With its stacked CMOS sensor technology

with a compact form factor, USB 3.2 10Gbs data

developed by Sony, the E9’s square pixel array with

communication, and long camera lifetime.

3.76 µm pixels and high dynamic range (>80 dB) combine to create a sensitive, versatile, and costeffective low noise camera ready for integration or benchtop use.

“The Retiga E platform outperforms many scientific CMOS cameras available today. The Retiga E9, with smaller pixels, captures better resolution at low magnification. Additionally, the dynamic range

With a combination of low readout noise, high

of Retiga E cameras is better than most scientific

quantum efficiency, small pixel size, and large full

CMOS cameras, even when compared to those

well capacity the Retiga E9 is an ideal solution for

with larger pixels.

imaging low-light signals for the extended periods of

cameras are limited to seconds of exposure due to

time required in gel documentation, DNA and RNA

dark current, not the tens of minutes possible with

sequencing, qPCR, fluorescence/phosphorescence

the Retiga E9. This makes the Retiga E9 one of the

imaging, and general microscopy applications. The

most flexible and powerful cooled CMOS cameras

9-megapixel, 3k x 3k array matches the field of view

available today,” states Product Manager Phil Allen,

of many scientific imaging instruments, microscopes,

PhD.

and other C-mount-based optical systems.

www.teledyne.com

And most scientific CMOS

71

NEW PRODUCTS

The DP75 Digital Microscope Camera Empowers Microscopic Discoveries and Inspections The DP75 digital microscope camera breaks new ground in microscopic imaging. Users can capture brightfield or wide-wavelength fluorescence images using a single camera, eliminating the inconvenience of switching between multiple cameras.

One Camera, Multiple Applications Designed for a variety of challenging life science and industrial applications, the DP75 camera empowers researchers and inspectors to capture highresolution images from brightfield to fluorescence, even in the near-infrared (NIR) range.This innovative camera simplifies the microscopy imaging process, enabling users to concentrate more on their work.

Sharper Images, Clearer Insights The high-sensitivity cooled CMOS sensor makes it easier than ever to capture sharp, low-noise fluorescence images. With a fast frame rate of 60 fps at full HD and 22 fps at over 4K resolution, the camera provides smooth, fast, live images for easy framing and comfortable live observation.

Vivid Color from Cells to Surfaces Whether inspecting the intricate structures of a cell or the surface of an industrial material, the DP75 camera provides outstanding image quality and ease of use. The camera’s advanced multiple-axis color correction technology helps ensure true-to-life color reproduction, making your images as vivid as looking through the microscope oculars. In addition, its smart AI-based scene detection automatically recognizes the observation method and adjusts the appropriate imaging parameters, which allows for easy capture of high-quality images, no matter the observation method.

Qualitative Analysis Meets Intelligent Features Quantitative data collection is easy with the DP75 camera’s linear mode, and it enables a relative intensity evaluation of fluorescence expressions on a sample. Moreover, the camera enables you to easily overlay fluorescence and brightfield images

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ISSUE 73 MARCH 2024

with pixel precision so that you can precisely identify the locations of fluorescent expression with the morphology of your specimen. For industrial applications, the camera’s live high dynamic range (HDR) feature provides high-fidelity images, capturing textures, flaws and previously undetectable defects with outstanding clarity.

Flexible and Efficient The DP75 camera is both powerful and flexible.

With compatibility via USB 3.1 Gen2, this camera works with most PCs, making it an ideal upgrade for any existing system. Its wide field of view capabilities further ensure fast and efficient research and inspections. www.olympus-lifescience.com/camera/ color/dp75/

High accuracy sample positioner now offers up to 800 µm travel Queensgate’s NanoScan SP800 nanopositioning

SP800 has the fastest step settle and recovery

sample scanner offers high-accuracy positioning

time between Z stacks to offer market-leading

performance for longer ranges.

resolution.

Queensgate has added a new model to its range of

The SP800 has a slim profile to enable easy access

piezo-driven sample positioners. The SP800 offers

for illumination of the sample area. A wide variety

800 µm of travel (closed loop) with a resolution

of sample insert plates are available, including full

of 1.2 nm and a step settle time of under 10 ms. It

microtiter plates, making it suitable for a range of

can cope with up to 500 g loads, with higher load

applications in life sciences and materials:

capacity available on request.

As with the SP400 and SP600 models, the SP800 is compatible with all Prior Scientific motorized microscope stages and components as well as other leading microscope brands.

Coupled with the Queensgate NPC-D-6110 digital controller, the system is easy to use and quick to set up using standard 0-10 V analog input and output.

The digital control technology features variable acceleration/deceleration algorithms which allow

• Optical sectioning producing 3D images • Live cell imaging • Autofocus systems for time-lapse imaging • High content screening • Surface analysis • Wafer inspection https://www.nanopositioning.com/ product/nanoscan-sp400-sp600-sp800piezo-sample-positioner-scanner

exceptionally fine control of stage movements.

Thanks to this, the NanoScan SP400/SP600/

If you would like your new product information to appear on these pages, contact infocus Magazine at advertising@infocus.org.uk The announcements in this Section are compiled by the manufacturers. They in no way represent a recommendation by the Royal Microscopical Society for any particular instrument or equipment. The Royal Microscopical Society does not endorse, support, recommend or verify the information provided on these pages.

73

Reflections on the RMS Diploma: Study, Mentorship, and Oversight Tim Young, Joëlle Goulding, Susan Brooks

If you are an RMS Member using microscopy or cytometry as part of your career, you can apply to study for an RMS Diploma - a flexible, portfolio-based, international qualification designed to complement and fit around your current employment.

The Diploma is intended to be of a similar standard to a Masters degree. Projects are designed by the candidate with the assistance of their line-manager, and mentorship from existing Fellows of the Society. This approach ensures that the study is both challenging and rewarding whilst complementing the candidate’s existing employment.

Here, we hear three different perspectives on the RMS Diploma, from some of those currently involved in the initiative.

Tim Young: Balancing an RMS diploma with full-time work

carry on developing my project, without reducing

Current Diploma student Tim Young is

my paid hours and for a fraction of the price of a

a Laboratory Supervisor for the Lehner

Masters degree. Furthermore, with my son starting

Group, CITIID, Jeffery Cheah Biomedical

school soon, the flexibility offered by the diploma

Centre, Cambridge Biomedical Campus In 2021, whilst still under COVID restrictions and things were quiet in the lab, I had time to think, plan, experiment and write my own project whilst undertaking all of my regular laboratory management duties. I was reading microscopy papers alongside almost unlimited access to the unit’s Zeiss 880 LSM. I was able to make great progress whilst increasing my confocal competency and producing some satisfying images too.

When I read about the RMS diploma on their

74

website, it seemed too good to be true; I could

ISSUE 73 MARCH 2024

was perfect for this stage in my life.

However, this was not to last long; after six months my PI moved his lab to Germany and I was unable to follow. I found another position at Cambridge University, and agreed to join as their Lab Manager with the condition that I could continue my diploma. Although my manager is supportive of my studies, the balance between my paid duties and my diploma has been much more challenging to maintain. I would like to share some of my insights from these last two years for anyone considering an RMS diploma.

Tim Young

issues, as my workload, like many laboratories, is variable and can drastically increase at little or no warning.This summer, I made the difficult to decision to stop all diploma activities during a particularly busy period in the lab. It was the right decision as the alternative could make all situations worse.

Thankfully, the RMS team understand that diploma students are usually full-time workers, not full-time students and that there are external responsibilities.

Because of this they are realistic about expectations, and you should be too; if you are devoting a half-

The most crucial aspect to consider is passion.

Whilst you are supported during the diploma, ultimately you are the driving force behind it and without a passion for microscopy it is not worth pursuing.

Does your job and your life circumstances allow time for study? Passion is key but without time in your week to fit in some reading, writing and (ideally) microscopy it will be stressful and a struggle from day one. Most employers support professional development amongst their staff, especially in academia, and if you are considering a diploma, your Line Manager or PI should be your first stop. You may be able to negotiate some reduced duties or set aside half a day for dedication to your studies.

For me, it began with a casual conversation, but such topics can be taken to HR or senior management for more serious talks.

It is advisable to know exactly what you are asking for in advance and to remember to display the passion for it; you don’t want to meet resistance at the first meeting. Have a plan as to how you can fit the diploma around your current workload and how it can complement the lab or facility’s remit.

Although I negotiated to continue my diploma in my new role, it was under the condition that my managerial duties must always take priority. This may be helpful to emphasise to your employer as assurance that your regular duties will not be neglected. However, it does come with its own

day a week to your diploma, do not expect the same experimental output as the full-time PhD student working alongside you. This is why the RMS are generous and flexible with their diploma requirements allowing between two and five years with ‘regular’ circumstances.

A particular difficulty I have experienced is starting and maintaining longer-term experiments. My work with stem cells proved challenging due to the regular passaging requirements which I was unable to continually maintain due to lab management work taking priority.

Planning for such circumstances is difficult alongside a variable and uncertain workload. Here a supportive co-worker is invaluable: someone who can cover your work at short notice, just to keep that experiment going or the cells alive. If this isn’t available, then you may need to re-think your experimental plan. For example, factor in pauses where you can stop, as needed, and re-start when things settle down.

As the saying goes, ‘make hay while the sun shines’.

When your workload allows be prepared to re-start experiments quickly to fully utilise quieter periods.

Make meticulous notes detailing all experimental protocols and results. These proved invaluable to me after months of unrelated work.

Have clear objectives. These can be laid out and discussed at your progress meetings and should be reviewed regularly with your mentor.

Balancing work, study and life will be a challenge, so it would be foolish to pretend it won’t be. But if

75

you go in with realistic expectations of yourself, a supportive manager and most importantly, passion, you will give yourself the best chance.

Dr Joëlle Goulding: Being a Diploma Mentor Dr Joëlle Goulding (FRMS) is based at the Centre of Membrane Proteins and Receptors (COMPARE), University of Nottingham, UK. Over the last few years I have become heavily involved with mentoring, something I had never really considered before. Part of this was because I thought that I didn’t have enough experience to pass on. To me the benefit of having a mentor is clear – someone to bounce ideas off, gain advice over tricky decisions and to be a sounding board.

I didn’t have the confidence to offer this. What we often overlook, however, is how much experience - both in work and life - that we have amassed.

When I stepped back to think about it, I realised I have experienced a number of job roles, written dissertations and given talks, survived fixed-term contracts and maternity leave which then led on to juggling childcare with a career. I have also gained expertise in a number of different imaging modalities, worked up and troubleshot protocols, and pursued collaborative and independent research interests. In the end it is all this experience together that actually makes me a very useful mentor for a diploma student …. Hopefully!

I am involved in the running of both my faculty mentoring scheme at the University of Nottingham and also the mentoring scheme which is offered by the RMS. Whilst both schemes are largely focussed on career and professional development, their angles and method of working are slightly different.

And again the mentorship as part of the Diploma has its own slant. As a Diploma mentor your main purpose is to support the Diploma student through their passage of work. This does not mean directing their project as a supervisor would, or indeed, judging the work as an examiner. You are also the contact between the mentee and the organising body, the RMS. Whilst representing the RMS in this fashion, you still have an outside view on the programme of study and hopefully a bigger picture view on how the diploma in the long run could influence the student’s career.

My own role as a Diploma mentor has varied, this was my first time in this role and a bit of a steep learning curve into what was expected. This wasn’t an unpleasant curve but there is a level of responsibility about knowing what is expected of the student and guiding them in some of the course credits, such as the technical essay and identifying a suitable training course. For this, luckily, I could draw on my experience, and through working with the mentee, realise I DID have the experience to pass on. Hopefully my mentee hasn’t suffered, but if I take on the role again I will be a lot more prepared!

What I didn’t realise was the vested interest that I developed in their career, being keen to make them aware of opportunities, conferences, societies and options that I, in their place, would have found invaluable. Your relationship has been set-up purely for this period of study, however this is likely to be a connection, a friendship that will continue.

Overview: Professor Susan Brooks Susan, who is based at Oxford Brookes University,

is

Qualifications Joëlle Goulding.

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ISSUE 73 MARCH 2024

Chair

of

Committee

the

RMS

which

meets

twice a year to discuss current diploma candidates’ progress reports.

Individuals who undertake the RMS Diploma

expertise, a scientific meeting or conference, and

always report very positively on the experience.

also undertake an outreach or public engagement

When we have asked them “what has the Diploma

activity that promotes microscopy. The programme

done for you professionally and personally?” we

is designed to be challenging, rewarding and fitting

receive responses such as “(it has)..showcased my

with, and complementing, your existing employment.

enthusiasm for microscopy to future employers…

Your final portfolio will demonstrate a range of

(I’ve

the

skills and capabilities; not just technical, but also

community”; they say that it’s been “professionally

made)

invaluable

contacts

within

your ability to communicate these to both specialist

.. a great experience”, and that it has brought them

and non-specialist audiences, and your commitment

‘’professional recognition” and “credibility amongst

to continued professional development.

my peers”.

Undertaking the Diploma is therefore a clear

So, what is the Diploma, and who is it for? The

demonstration of your commitment to improving

Diploma is an international programme – we have

your skills and to promoting awareness of

Diploma candidates worldwide, including from

microscopy to a wider audience. You will improve

Germany, Switzerland, Portugal, Australia and New

your existing skills - and acquire new ones - that

Zealand – and, of course, from all parts of the UK.

will benefit your career and your employer for

They are based in Universities, research institutes,

many years to come. During your study, you will

industry and schools.

have the support of a dedicated RMS mentor. The

Typically, those undertaking a Diploma are working in a role where microscopy or cytometry is a major part of their employment and they wish to develop their professional competence, challenge themselves to learn new things, and develop additional skills. There is also the desire to receive formal, professional recognition for their expertise within their specialism. It is considered to be of

Society is currently pursuing a path to become licensed by the Science Council to award Chartered Scientist. When this is in place, a Diploma should provide a pathway to Chartered Scientist. As one of our Diploma candidates said: “(this was)….an incredible opportunity to advance in my career as a microscopist”.

Susan Brooks.

equivalent status to a masters level qualification.

The Diploma is a bespoke, portfolio-based qualification

that

flexibly

fits

around,

and

complements, your current professional role. During your Diploma programme, which takes a minimum of two years but can be extended (depending on your work commitments), you undertake a workplace-based practical project, based in your employment, and designed by yourself with the assistance of your line manager and with expert support from the RMS. You then present a report on your project and a technical essay demonstrating your

knowledge

and

understanding

of

the

microscopy / cytology techniques and approaches you have used. In addition, you must attend a course supporting you to further develop your practical

Find out more about the Diploma at http://www. rms.org.uk/study-read/rms-diploma.html/ or contact Kate Wooding kate@rms.org.uk to discuss your ideas for a Diploma project.

77

RMS Diploma: Meet our latest Candidates!

We are very pleased to welcome five new candidates onto the RMS Diploma programme, and wish them all the very best of luck as they progress their projects.

The RMS Diploma is a flexible, portfolio-based qualification designed to complement the candidate’s existing employment.

Projects are designed by the candidate, with assistance from their line-manager and input from existing Fellows of the Society.

Here is a run-down of our latest candidates, and their chosen projects: Samuel Davis, Loughborough University, UK

Scanning Electron and Focused Ion Beam (FIB) systems will be used to collect images, employing

Project: Electron Microscopy Study of Precipitates in Energy Generation Steels

a combination of modern detectors, and optimised

Using a multimodal approach to compare precipitate

currently referenced standards.

operating parameters for each system. These will be compared to benchmark examples cited in the literature with a view to improving upon the

imaging techniques of power generation steels. Both

Susan Duncan, John Innes Centre, UK Project: Development of Quantitative Single Molecule RNA Imaging in Plant Crop Species Quantitative single molecule RNA imaging is highly complementary to next generation RNA sequencing approaches. Although lower throughput, imaging has an advantage over sequencing as it reveals subcellular RNA localisation patterns that can have significant consequences for gene regulation.

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ISSUE 73 MARCH 2024

has. Using

confocal

and

super-resolution

microscopy, the localisation and dimerisation status of ATM will be observed before and after DNA damage in neuronal and non-neuronal cells, to observe its pattern of behaviour and speculate as to its significance in the DNA damage response.

Despite advances in RNA imaging in many other model organisms, for plant biology this approach is currently limited to a few tissues in the model plant Arabidopsis thaliana. The project proposed for this Diploma aims to expand this imaging approach to a wider range of tissues in commercially important crop plants including wheat, sugar beet and oil seed

Pamela Twist, Safi Biotherapeutics, UK Project: Application of Deep Learning Tools for Imaging Cytometry to Enable Label Free Characterisation of Manufactured Red Blood Cells

rape.

Rebecca Gascoyne, University of Birmingham, UK Project: Investigation of the dynamic localisation and dimerisation of ATM kinase Ataxia-Telangiectasia Mutated (ATM) is the central regulator of the DNA damage response in cells.ATM is mainly located in the nucleus or cytosol but has also been found at synapses – and the significance of this is unknown. It is known that ATM exists both in a monomeric and dimeric form but is unknown what significance the localisation and dimerisation

In the manufacture of red blood cells (RBCs), it is crucial to track the maturation of cells in the bioreactor from a hematopoietic progenitor cell to a mature RBC. This can be achieved by fluorescent staining of maturation markers. Using imaging cytometry, we can collect the fluorescent signal of cells along with their corresponding brightfield image that captures morphological changes of maturing RBCs. My study aims to investigate if we can apply deep learning tools to our imaging cytometry data, so we can characterise the stage of maturation of manufactured RBCs in a label-free manner.

79

Krunoslav Vinicki, Digicyte Ltd, Croatia Project: Rapid and NonDestructive Analysis of fresh or Formalin-Fixed Tumor Biopsies Using Fluorescence Microscopy and Artificial Intelligence for Image Reconstruction The study aims to develop and validate a new approach in microscopy for the rapid and non-destructive analysis of fresh tumor biopsy samples.With this method I’m planning to address a longstanding need in pathology for real-time, on-site biopsy examination while preserving the tissue for further analysis. This could expedite the initiation of targeted therapy and reduce the need for repeat sampling due to insufficient material.

European Light Microscopy Initiative 2024 (elmi2024) 4 - 7 June 2024, Liverpool, UK The event will encompass: • a blockbuster meeting programme covering all the latest techniques, applications and technology. Topics include: New Technologies, Imaging Across Scales, Super-resolution and Nanoscale Imaging, The AI Revolution, The Science of Tomorrow Today, and Multimodal Imaging; • a wide range of companies showcasing their latest technology and running workshops timetabled outside of the main meeting programme;

ACC Convention Centre and Exhibition Centre, Image Credit: Ant Clausen

Event Management by

• an accompanying exhibition in the purpose-built hall alongside posters, food and drink; • a community workshop space at the heart of the exhibition, with many groups hosting meetings and running workshops; • an event dinner with networking at the Rum Warehouse.

Find out more at www.elmi2024.org

Royal Microscopical Society 37/38 St Clements Oxford, OX4 1AJ www.rms.org.uk Registered Charity: 241990

royalmicrosoc #elmi2024

R E P O RT

Materials Research Society Fall Meeting 2023 Boston, Massachusetts, USA 27 November – 1 December

T

he annual Materials Research Society (MRS) Fall conference was held in Boston at the Hynes Convention Centre from 27 November to 1 December 2023.

As the plane from Dublin descended into Logan airport and the Prudential Centre came into clear view, the excitement for the upcoming MRS conference took over! As one of the largest materials science conferences in the world, with regular attendances of over 5000 people, it was sure to be packed with amazing talks and posters.

Given the recent funding rounds on both side of the Atlantic for more research into the topic, it was obviously clear that the theme of this year, at least in the physical sciences, was quantum-based materials and devices. As a microscopist (and with this being an in-house microscopy journal), there was something for everyone here. I attended the EELS and 4DSTEM sessions, and was inspired by the cutting-edge materials microscopy being performed, especially on quantum phenomena and materials such as the Rashba effect and topological insulators.

The conference also had a series of excellent plenary talks which I had the privilege of attending.

These included the recent Nobel Prize in Chemistry winner, Moungi Bawendi of MIT, and Takashi Taniguchi of NIMS, Japan - the go-to guy for excellent quality hBN crystals. It was fascinating to see their academic career journey and helped to emphasise the point that since there is no easy, straight-line path to success, you should make your own path!

I presented my works on “Structural Characterisation of BaZrS(3-y)Sey Perovskite Thin Films via Scanning Transmission Electron

Me inside the Hynes Convention Centre with the big MRS sign.

Microscopy” and “Thermoelectric StructureProperty Relationship Establishment in TlGaSe2” as oral presentations at the end of the week. Despite being tired from such a long but engaging week, the talks went very well, and I had a number of interesting discussions with people from various fields in materials science about my work, which was quite rewarding!

As I am within the last few weeks of my PhD, MRS Fall 2023 was my last conference as a PhD student, and I am delighted that it was! Having spent some time in MIT last year, it was lovely to catch up with old mates and colleagues, as well as make new ones. I am very grateful to the RMS for the financial assistance to enable me to attend this conference.

Tigran Simonian School of Chemistry & Advanced Microscopy Lab, Trinity College Dublin, Ireland

81

From the archives: A ‘double-take’ from 1989 Many long-standing RMS members will recall that 1989 was a celebratory year for the Society, marking 150 years since its inception. Later this year, we’ll be looking back at some of the anniversary events that took place – as documented by The Proceedings – but for now, our attentions turn to an unusual pair of photographs from Volume 24 of the Proceedings, published in January 1989.

The issue includes a list of speakers due to give talks later that year, at – as it happens – a special meeting to mark the aforementioned anniversary. Those speakers included Dr Hans Tanke, of the University of Leiden, Netherlands. However, his biography photo appears in double, giving - at first glance - the impression of an unlikely production error.

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This was no error, but rather, an inventive piece of editing from Proceedings editor, Peter Evennett.

(You can read a fascinating interview with Peter, which appeared in our June 2021 issue.)

As it happens, the presentation of the photographs was linked to Dr Tanke’s decision to submit a pair of images, apparently

lecture

taken at a photo booth. A footnote beneath Dr Tanke’s listing directs readers to a later page to seek the explanation (see image above).

To round the story off, infocus managed to get in touch with (the now) Professor Tanke, who retired as Head of Department at Leiden in 2017. He was kind enough to share some memories of his involvement in the 1989 anniversary events – and, it turns out, he even remembers the photographs

he threw a handful of matches on the wooden floor and invited the participants on the first row to count the number of crossings with the structure of the wooden floor: of course his prediction of the outcome was right!”

On the subject of the photographs, he added: “Indeed, they were taken at a kiosk, since the real digital world was a little ahead. I have never realised though, that they represent stereo pairs. My wife - whom I have been married to for 48 years liked them, and said

from 35 years ago!

“in those days, you

He said: “I do remember the 150th celebration of

looked much more......

the RMS. It was a privilege to lecture there instead

(you may fill it in!).”

of Professor Johan (Bas) Ploem, former president of

…Well,

the RMS, who was originally invited. I remember the famous lecture theatre, in which I was told, Isaac

lovely, recent photo

Newton had presented his work years ago.

of Professor Tanke to

“The speaker after me was DrWeibel, who explained his theory of stereologic measurements. During his

whatever

she meant, here’s a

bring us up to date: Hans Tanke in a recent photo.

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Submission Guidelines

infocus is the Royal Microscopical Society’s (RMS) vibrant and striking quarterly magazine for members. It provides a common forum for scientists & technologists who use any form of microscope, including all branches of microscopy. Published four times a year, infocus is free to members of the RMS. infocus features articles on microscopy related topics, techniques and developments, an events calendar, news, event reports, book reviews, new product information, and much more. infocus welcomes submissions of: Articles - Full articles or reviews of general interest to microscopists, of approximately 30004000 words (excluding references), with images/ figures (as many as appropriate, 4-8 as a guide).

Longer articles can also be considered.

Short Articles - Short topical articles, review articles or articles providing hands-on help for microscopy methods.

Primer Articles - Short general articles that are focussed on specific techniques.

Debuts - Student articles publishing emerging results from a project. Results may still be incomplete, but areas of progress/problems should be highlighted, with the aim of provoking feedback.

Book Reviews – if you are a member of the RMS and are interested in writing book reviews for infocus, please contact Owen Morton owen@rms.org.uk.

Please see recent issues of infocus for examples of articles and reviews. To request a sample copy of infocus contact owen@rms.org.uk If you are interested in submitting to infocus, contact: editor@infocus.org.ukj

Article Text

• Abbreviations should be used sparingly and only if a lengthy name/expression is repeated throughout the article. When used, the abbreviated name or expression should be cited in full at first usage, followed by the abbreviation in parentheses.

• Authors should provide a photograph, brief biography as well as contact information that will be published.

References

References in the text should be in the form Joy (2000) or Joy & Williams (2000). For three or more authors, use the form Echlin et al. (2000). The reference list should: • be listed in alphabetical order of first authors’ surnames.

• (where a journal is cited) - include authors’ surnames and initials, date of publication, title of paper, name of journal, volume number, and first and last page numbers.

• (where a book is cited) - include authors’ surnames and initials, title of book, year of publication, edition, followed by publisher and town, county/state (and country if necessary) of publication.

• (where a URL is cited) – include authors’ surnames and initials, year of publication, title of page, URL and date accessed.

Images / Figures • Figures can be one column/half page width, 65.5 mm or two column/full page width, 135 mm.

• Larger images may fill the page/spread. A full page of the magazine is 170 x 250 mm, a double page is 340 x 250 mm.

• Text in figures (labels, axis labels, legends, etc) should be Helvetica or Arial, 8pt size.

• Figure and table captions should be listed numerically at the end of the article text

• Text should be in a standard font (e.g. Times New Roman or Arial) at a size of 12 pt.

• Line weights and line strokes should have a maximum value of 1 and a minimum of 0.25.

• Articles should begin with a brief summary, which accurately summarises the content and is intelligible without reference to the text.

• For graphs and plots, whenever possible, please submit vectorized images.

• As much as possible, please avoid white spaces.

• Footnotes and appendices should not be used unless absolutely necessary.

• All images must be high resolution – 300dpi or more.

• The hierarchy of headings within the text should be clear.

• Submission files should be in CMYK format and can be supplied as tiff, jpeg or eps files.

• Spelling should conform with The Concise Oxford Dictionary and SI units must be used.

• Images MUST include scale bars or field widths where relevant.

Double page of magazine, 340 x 250mm (Trim size) 84

ISSUE 73 MARCH 2024

• Total number of images/figures/tables should not exceed 15 including tables.

Proofs

Prior to publication, authors will be sent a PDF of the article by email for approval.

Authors should ensure articles are thoroughly checked before submission – proof amendments should be limited to minor corrections only.

One column/half page width, 65.5mm Figure 1. Width of figure or table confined to one column.

Offprints Five hard copies of the issue in which the article is published will be sent to the author, together with an emailed PDF of the article.

Copyright Authors are requested to assign copyright to the RMS. However, authors may make copies of their own articles without seeking permission from the RMS, provided that such copies are for free distribution only (they must not be sold) and provided that infocus is properly acknowledged (issue number, month and page number should be given). Permission to reproduce material from infocus in other publications will not be given to third parties except with the consent of the authors concerned.

Authors are responsible for obtaining permission to reproduce copyright material from other sources.

Approval for reproduction/modification of any material (including figures and tables) published elsewhere should be obtained by the authors before submission of the manuscript and the source of the material should be properly acknowledged. Authors are responsible for any copyright fee involved.

Authors are requested to complete and submit a signed copy of our copyright sign-off form. This is available on the RMS website (www.infocus.org.uk).

Two column/full page width, 135mm

Figure 2. Width of figure or table spanning full width of page.

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