infocus Magazine - Issue 69, March 2023

Bugs up Close RMS Summer Studentship Reports 2022 mmc2023: Register now!

The Expansion Microscopy User Group: Cultivating a new scientific community

Breaking Barriers with Today’s Most Powerful Workhorse S/TEM

The F200 does more to change the accepted views of uncorrected S/TEM than ever before. Its enhanced cold field-emission source, large-area silicon drift detectors, and electron optics stability are the foundation for capabilities that exceed the expected.

• Atomic-scale imaging – even for thick specimens

• High spatial resolution – even at low kV

• Spatially-resolved analytical information with EDS/EELS

• Optimum Bright Field (OBF) STEM – Maximum S/N at low dose

• Light element detection

With the F2, you’ll never have to compromise. Get the performance you need in one powerful TEM that does more than you’d expect.

features

4 Bugs up Close

John Hutchison, Hon FRMS

28 RMS Summer Studentship Reports 2022

Ben Watson, Catherine Read, Hale-Seda Radoykova

50 mmc2023: Register now!

58 The Expansion Microscopy User Group: Cultivating a new scientific community

Owen Morton

and other features

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.

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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

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ISSN: 1750-4740

© 2023 Royal Microscopical Society

infocus is published four times per year by the RMS. Designed and produced by The ImageWorks. 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.

FROM THE SCIENTIFIC EDITOR

Dear Readers,

Happy New Year! I hope you all had a relaxing break over the festive season, enjoying time with family and friends (and pets!) – and also that the first months of 2023 have treated you well.

It is with great pleasure that we bring to you our first infocus of 2023, and our first, fully digital issue! I really hope you enjoy all our online content, as we embark on this exciting new voyage of discovery.

Speaking of exciting discoveries, RMS History Committee Chair John Hutchison describes in this issue how a box-full of beautiful slides dating back to the 19th century ended up in the RMS archive. John provides a great description of how slides were prepared back then – and if you want to take a closer look, the slides will be on display at mmc2023 in Manchester, as part of the famous RMS Learning Zone. mmc is the flagship microscopy event of the Society and this year we are all going to be there fully in person.

Every year the RMS awards summer studentships for students in their second year of study (or third year in a four-year degree), giving them the opportunity to carry out projects involving microscopy or image analysis in physical or biological sciences. It is a great way to foster a new generation of microscopists. The awardees write a report at the end of the project, and we are delighted to publish these in this issue. Hale-Seda Radoykova, a student at King’s College London, talks about her project on designing a pipeline to analyse fluorescent labelled microtubules of yeast; Catherine Read, of the University of York, describes her project on analysing and imaging liquid droplets to study liquid-phase separation of CO2-fixing enzyme; and Ben Watson, a second year student at University of Strathclyde, describes his work on 3D printed optical elements, characterising and quantifying their surface quality.

I will be in Manchester for mmc2013, so if you would like to talk about infocus, and how you can contribute to the magazine, I will be more than happy to sit down for a chat. I hope we can all meet again – either at mmc or at other meetings and congresses throughout the year.

Slàinte!

Leandro Lemgruber

Leandro Lemgruber

COVER IMAGE: Battery electrode precursor powder

This image, by Lucia Spasevski and Alexandra Stavropoulou, is an SEM-EDS chemical analysis of battery electrode precursor powder. Equipment used: Gemini 460 and Oxford Instruments Ultim Extreme windowless EDS detector. Magnification x5300.

Bugs Closeup

Dr John Hutchison Hon FRMS

During the early days of the RMS, its (mainly wealthy) gentlemen, since women weren’t allowed to attend, would meet regularly, when members would discuss their latest microscopes and share some of their observations, which would most likely be hand-drawn at the time. Obtaining interesting specimens was more of a challenge however, as many of the members had neither the skill nor time to spend mounting slides with suitable materials. The Victorian fashion of examining novel objects was also an influence behind the appearance of professional slide mounters in the mid-1800s who, recognising a potentially large market opportunity, began to prepare slides of a wide variety of objects. These slides, mainly 3 x 1 inches in size following the adoption of the (still widely used today) ‘RMS standard’, have survived in large numbers and provide a fascinating insight into the early days of ‘popular’ microscopy, and of the RMS.

We recently received a generous gift of a cabinet of antique slides, which was accompanied by an unusual, folding, Swift microscope, which dates back to the early 20th C.

There are over 800 slides in all, some dating back to the 1860s and are in several categories: entomology, botany, histology, petrology, and diatoms – the salicaceous (glass) coatings of algae micro-organisms commonly found in water (both fresh and salt). These were collected, cleaned and mounted, often in very artistic arrays and will be described in a future article.

“Wow!” slides

Several slides have a very obvious “wow” factor. These include whole insects, carefully laid out and preserved in resin (usually Canada Balsam) between slide and coverglass.

An additional eight boxes of slides were also included. The donor, Nigel Hill, came across them while going through his late father’s belongings. He had been given them by “a friend of a friend”, who thought that the young Nigel might be interested in biology or botany. Nigel, however, preferred sport and then studied languages before a career in the world of banking. The microscope and slides were thus wrapped up in newspaper and put in the attic. The newspaper was dated 19th May 1967 and was only uncovered in the summer of 2022 – some 55 years later!

Figure. 1 Cabinet containing 20 trays of antique microscope slides.

Figure. 3 Dragonfly nymph. Note the extended pincers.

Figure. 2. Selection of “whole insect” slides from the late 19th to early 20th Century.

The dragonfly nymph, remarkably well preserved here, was a voracious predator in the ponds where it would spend several years before transforming into the free-flying adult insect. Figure 3 clearly shows the formidable pincers, which could be extended – as shown here – to draw the victim back into the nymph’s jaws.

A mature dragonfly is shown in Figure 4 for comparison.

For many years the Colorado beetle has devastated potato crops in the USA and Canada. Fearful of it attacking British crops, what was then the Ministry

of Agriculture ran a campaign in the 1960s (Figure 5) warning of the dangers of these beetles, and in particular their larvae which would devour the leaves of the potato plants, thereby destroying crops. Another of our “wow” slides (Figure 6) shows one of these larvae, which is only ~ 15 mm in length.

Frederick Enoch, possibly the most famous slidemounter of his time, was particularly well-known for his ‘whole insect’ slides, and several in the collection illustrate the superb layouts he created as seen in Figure 7, which shows a cranefly (aka ‘daddy long-legs’).

The example shown in Figure 8 is a fungus gnat. This slide includes a label confirming the preparer as Enoch and the address allows us to date the slide to 1882, when he lived there.

Figure. 4. Mature dragonfly.

Figure. 5. Poster campaign for Colorado beetle crisis.

Figure. 6.Colorado beetle larva.

The larger, whole insect slides would often have the specimen’s soft innards carefully removed by soaking in potassium hydroxide before cleaning

spacer ring and small, glass cover slip was used to mount smaller insects or selected parts in a liquid while preserving their solid shape, as opposed to

and drying. The exoskeleton was then laid out in a resin or balsam on a slide before covering and sealing with the cover glass. This was an elaborate procedure and well-mounted examples survive that date back to the late 1800s.

The development of ‘closed cell’ slides, with a

the flattened exoskeletons of the ‘whole insects’ shown above. Many such mounts were sold by the London firm Clarke & Page, including some supplied by other makers such as Enoch. Many of their slides include “prepared without pressure” on their labels and preserve the specimen in great detail. An example of a whole insect in a closed cell is shown in Fig. 9. The snipe fly was named after the snipe, as its proboscis was thought to resemble the downward pointing bill of the bird!

Separate parts of insects were also mounted in large numbers, some of which show astonishing details, such as a blow-fly proboscis, shown in Figures 10 and 11.

Figure. 7. Cranefly.

Figure. 8. Fungus gnat.

Fig. 9. Closed cell mount containing a snipe fly. Note that the embedding medium has discoloured over the years.

Figure 12 shows a rather startling cranefly’s head, contrasting with the whole-insect mount shown in Figure 7.

Figure 13 is an example of a mount with a black background which would highlight an opaque specimen when viewed with incident light. It shows a closed cell containing a diamond beetle, commonly found across Europe. This one is some 7mm in length.

Wings of similar beetles along with butterfly scales and diatoms were also used to make elaborate arrangements, which were popular in the late 1800s

and into the early 1900s. Figure 14 is an example, created by William Firth, one of the foremost diatom mounters of his time. His mounts are still keenly sought-after today.

Parasites

These are organisms that feed off a host animal or insect, and in this section several examples will be described.

Blood-sucking ticks are a major problem for any sheep farmer and require the sheep to be immersed in highly toxic fluid to eliminate them. They also carry Lyme’s disease, and their bites can cause serious illness in humans. Figures 15, 16 and 17 show an example of this bug, about 5 mm long. The sharp claws at the end of its forelegs (Figure 17) would provide a strong grip on its prey. The monogram “JB” on the paper cover of this slide (Figure 15) indicates that it was produced by John Barnett, a mounter who was active in the 1870s.

Figure. 10. Blowfly mouth and proboscis

Figure. 11 Details of blowfly proboscis.

Figure. 12 Head of cranefly.

Figure. 13 Diamond beetle in a closed cell mount.

Figure. 14 Arrangement of beetle wings, butterfly scales and diatoms.

Birds are also affected by parasites, and Figure 18 illustrates a blood-sucking louse (anopluran) that was found on a sea eagle. They are a common parasite among large birds of prey.

At the other end of the scale is the bee louse, which attacks honeybees. This tiny, wingless arthropod is only ~ 1mm in size, but can cause havoc when a colony of bees is infested. Figure 20 shows an example. To hitch a ride on their honeybee hosts, these lice need a truly phenomenal grasp. Now a recent study reveals how they manage to walk around on a flying bee while exhibiting what researchers say is the highest attachment force per body weight of any land-based insect ever measured.

Our largest bird, the ostrich, is not above such attacks by these tiny creatures, and Figure 19 shows an ostrich louse ~ 4 mm in length that causes serious problems in these birds. Ostriches are farmed for their meat and their skin is used for leather which, believe it or not, is used in ‘designer’ handbags (!). Infestations of these lice can not only damage feathers, but produce severe pitting in the leather, rendering it unusable.

Figure. 15. Mount by John Barnett containing a sheep tick. Note the “JB” monogram.

Figure. 16 Whole sheep tick, ~5 mm in size.

Figure. 17. Claws on foreleg of sheep tick.

Figure. 18. Louse from a sea eagle.

Figure. 19 Louse from ostrich. Note the claws on the forelegs.

This force relies on the parasite’s highly adapted feet, called ‘tarsi’, which are equipped with toothed claws. Each foot has a total of 28 teeth, or claw tips, which let the parasite lock onto sparse honeybee hairs during flight.

“The claws are unique, from what we know so far. Usually, insects have claws with one tip only. A few species have two to three tips. But this species possesses comb-like claws with several tips and deep interstices [gaps],” says Thies Büscher, a zoologist at Kiel University.

provides a rich resource for anyone seeking to explore the early days of the RMS and what our early members looked at with their microscopes.

Another of our members, Stuart Clague produced an article for infocus describing some of his own private collection of slides (see issue 48, December 2017). A more scholarly work by the late Brian Bracegirdle, a former Executive Hon. Secretary of the RMS, was published by the Quekett Microscopical Club in 1998. This volume contains details of most of the slide preparers of the Victorian and Edwardian times and shows many examples of their work. A copy of this publication is in the RMS library. There are also numerous collectors’ websites which provide more information on antique slides.

Of course, any discussion of blood-suckers must include fleas, which are widespread throughout the animal kingdom, and we conclude this report with images of these pests in Figures 21 (a) and (b).

No description of flea images could fail to mention Robert Hooke’s truly remarkable picture, published in his Micrographia in 1665.This wonderful etching is shown in Figure 22.

Conclusion

This short article aims to provide a fascinating glimpse into the small world that was revealed by microscopy in the late 19th C and early 20th C. Insects would appear to leap out of their balsam cages, tiny bugs appeared in great detail, and people’s curiosity for seeing new things led to an upsurge in microscopy (by now not confined only to wealthy gentlemen), particularly as ready-prepared slides or mounts became widely available. Our slide collection

I am pleased to reveal that we are planning to bring a selection of these antique slides to the ever-popular Learning Zone at mmc2023, the RMS’s flagship event. Visitors will be able to examine them in detail, using a microscope which dates from the same period. If you are planning to attend, please do make time to visit the Learning Zone and take a look!

Dr John Hutchison, Hon FRMS

Figure. 20. Bee louse.

Figure. 21 Examples of fleas from (a) boar and (b) mole.

Figure. 22 Robert Hooke’s iconic flea etching from 1665.

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.

Our online calendar includes all the details about forthcoming talks in the International Microscopy Focus Lecture Series – a joint, online initiative established between the RMS, and a number of international societies.

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

2023

March

6 – 9 Virtual Flow Cytometry Data Analysis Spring Course 2023 (Online)

8 Virtual International Microscopy Lecture Series - Professor Sumio Iijima (Online, multi-society event)

April

2 – 6 Botanical Microscopy Meeting 2023 –Norwich, UK (RMS-sponsored event)

3 – 6 Microscopy of Semi-Conducting Materials Conference - Cambridge, UK

17 – 21 Spring School in Electron Microscopy 2023 – Leeds, UK

18 – 19 Electron Backscatter Diffraction Meeting (EBSD) 2023 – Leeds, UK

July

4 – 6 mmc2023: Microscience Microcopy Congress 2023 (incorporating EMAG 2023) – Manchester, UK

7 Super-resolution workshop – Leeds, UK

10 – 12 Light Microscopy Summer School 2023, York, UK

13 – 14 Getting the most from your Confocal Course 2023, York, UK

23 – 27 Microscopy and Microanalysis 2023 (M&M 2023), Minneapolis MN, USA (Non-RMS Event)

September

5 – 8

Adhesion and migration in disease: Translational and therapeutic opportunities –Warwick, UK

November

16

flowcytometryUK 2023, Cambridge, UK

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

Featured RMS events

Botanical Microscopy Meeting 2023

2 – 6 April, Norwich, UK

Scientific organisers: Kim Findlay, Christine Faulkner and Richard Smith, John Innes Centre; John Hussey, University of Durham; Ulla Neumann, Max Planck Institute for Plant Breeding Research; Imogen Sparkes, University of Bristol

The 12th Botanical Microscopy Meeting is the latest in a long running series of RMS sponsored meetings, dating back to the 1960s, encompassing all aspects of bioimaging relating to modern plant cell biology.There will be a plenary speaker on the Sunday evening plus a further 7 keynote speakers, covering six scientific sessions. The rest of the programme will be chosen from offered talks, with a poster session on the Monday evening. Topics will be a mix of state-of-the art microscopy combined with the latest developments in plant cell biology, including morphogenesis, plant

membranes, organelle dynamics, plant-microbe interactions as well as quantitative imaging and image analysis. There will be an exhibition on Monday to Wednesday. The conference dinner will be at The Assembly House, a Georgian gem, described by historians as one of the most important buildings in Norwich.

Optional tours will be available around the JIC Bioimaging unit (light and electron microscopy facilities) and also to see The JIC Special Collection; this rare book collection is regarded as particularly outstanding, not only because it embraces five centuries of botanical literature, but also because it houses many works that are landmarks in the history of plant science, the earliest of which is Ortus Sanitatus, a herbal dated 1511. It also includes extremely beautifully illustrated and highly valuable works such Les liliacées by Pierre-Joseph Redouté, 1807.

We look forward to welcoming you to Norwich!

4 – 6 July, Manchester, UK

Make sure your diary includes mmc2023! One of the biggest events of its kind in Europe, mmc2023 will bring you the very best in microscopy, imaging and cytometry from across the globe. With six parallel conference sessions, a worldclass exhibition, workshops, satellite meetings, an international Imaging Competition and more, it’s simply the place to be for anyone who uses a microscope for work, study or pleasure.

Back at the superb Manchester Central conference centre for the first time since 2019,

the event will be taking place fully in person, providing the perfect opportunity for the scientific community to come together, make connections and share their research. As always, many of the leading companies in microscopy and imaging will be on hand to demonstrate the very latest equipment and technology at the exhibition, which is returning to Manchester from 4-6 July.

As with the previous events, you can expect a huge and varied scientific conference alongside Europe’s largest free microscopy and imaging exhibition filled with a huge number of free training workshops.

mmc2023 (incorporating EMAG 2023)

Light Microscopy Summer School 2023

10 – 12 July, York, UK

Scientific organiser: Peter O’Toole, RMS Vice President, University of York

The Light Microscopy Summer School is an annual three 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

Getting the Most From Your Confocal Course 2023

13 - 14 July, York, UK

Scientific organiser: Peter O’Toole, RMS Vice President, 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

presentations, the 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.

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.

image courtesy of Steve

17th Methods and Applications in Fluorescence Conference (MAF2022)

11 – 14 September Gothenburg, Sweden

The 17th Methods and Applications in Fluorescence (MAF2022) was held at Chalmers University of Technology in Göteborg, Sweden on September 1114th.

Initially scheduled for 2020, the organisers Marcus Wilhelmsson and Bo Albinsson warmly greeted the 350+ attendees before the meeting was kicked off in style by a keynote lecture from Stefan Hell, giving us an update on the progression of MINFLUX towards its new offshoot MINSTED.

These super-resolution techniques blow the mind with the achieved resolution of 1-3 nanometres as well as being photon-savvy - the high photon desire that is required for single molecule localisation microscopy, is delivered by the excitation laser and not the tracked target.

The following three days of the conference consisted of two parallel sessions per day with a further four keynotes, 10 invited talks and 10 oral sessions (talks selected from submitted abstracts). On the final day the MAF prize was awarded to Enrico Gratton in recognition of his contribution

within the field. All this taking place in the impressive lecture theatres of the Chalmers conference centre hosting incredibly comfy seating.

The conference attracted biologists, chemists, physicists and microscopists alike, bringing together a throng of scientists eager to share ideas, learn new concepts and to finally mingle again after too long stuck in a virtual meeting world. There was a definite vibrancy in the air as colleagues and friends joyfully greeted each other during the welcome event. Being a newcomer to this conference and knowing very few attendees, initially I was able to spot the few I recognised from twitter and finally meet a few scientists whose work I have read and admired in publications.

The two themes that jumped out at me and ran through the conference were nanoparticles and molecular rotors. Both concepts popped up in several talks, including new methods of creating nanoparticles, making them brighter and playing with the photophysics to turn on/off their fluorescence so these properties could be lent to advanced microscopy concepts. Molecular rotors were first introduced in the second Keynote by

Marina Kuimova of Imperial College London, who introduced us to viscosity sensitive molecular rotors that report on their environment through modulation of their fluorescence lifetime.

Variations of orientation sensitive molecules were also explored by Kai Johnsson, Max Planck Institute, Andrey Klymchenko, Université de Strasbourg, and Blaise Dumat, Ecole Normale Supérieure.This was a great forum in which to learn about new techniques and make collaborative links through which to explore and utilise these tools within our research.

I gave my selected talk, entitled ‘Ligand-Directed Fluorescent Labelling for the study of Membrane Proteins by Fluorescence Correlation Spectroscopy’, on the 13th September in the massive RunAn lecture hall.

Despite being especially nervous - it’s been so long for all of us speaking in person in front of others - it was fantastic to tell everyone about our new probes and the applications we’ve been developing to allow single receptor pharmacology to be studied. Alongside the lecture content were two days of 200+ posters and it was a pleasure to see the dedicated sessions so heavily attended. I was able to

search out those exploring fluorescence correlation spectroscopy and had several conversations bemoaning the joy and frustration associated with the technique!

The conference dinner was held at a hotel in the centre of Göteborg, where we were greeted by bubbly, a band and triumphant trumpetry from organiser Bo. A delicious, three-course meal was served with intermittent entertainment from a quad of singers and their guitarist, performing a medley of popular songs including, of course, Abba! The evening was a success with great discussions carried over from the scientific content and promises to look forward to the next scheduled MAF2024 planned for Valencia.

I would like to thank the Royal Microscopical Society for supporting me in attending this conference alongside funding from the Medical Research Council that supports my research.

Dr Joelle Goulding Research Fellow in Advanced Microscopy Centre of Membrane Proteins and Receptors School of Life Sciences University of Nottingham

Microscopy Journal of

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:

The structure of the eggshell and eggshell membranes of Crocodylus niloticus

A V Lensink, G E Swan, J G Myburg

The macro- and microstructure, elemental composition, and crystallographic characteristics of the eggshell and eggshell membranes of the Crocodylus niloticus egg was investigated using optical and electron microscopy, energy-dispersive X-ray spectroscopy (EDS), electron back scatter diffraction (EBSD) and computerised tomography. The translucent ellipsoid egg is composed of two basic layers, the outer calcified layer referred to as the shell and an inner organic fiber layer, referred to as the shell membrane. The outer inorganic calcite shell is further divided into an external, palisade and mammillary layers with pore channels traversing the shell. The external layer is a thin layer of amorphous calcium and phosphorus, the underlying palisade layer consist of irregular wedge-shaped crystals composed calcite with traces of magnesium, sodium, sulphur, and phosphorus.

The crystals are mostly elongated, orientated perpendicular to the shell surface ending in cone shaped knobs which forms the inner

mammillary layer. The elemental composition of the mammillae is like that of the palisade layer, but the crystal structure is much smaller and orientated randomly. The highest number of mammillae and shell pores are found at the equator of the egg, becoming fewer towards the egg poles. The shell thickness follows the same pattern, with the thickest area located at the equator. The eggshell membrane located right beneath and embedded in the mammillary layer of the shell; it is made up of unorganised fibre sheets roughly orientated at right angles to one another. Individual fibres consist of numerous smaller fibrils forming open channels that runs longitudinally through the fibre.

The basic measurements, and microscopic and chemical structure of the Nile crocodile egg was studied using several microscopy and analytical techniques. It was found that the translucent white egg has two basic layers; the outer shell and inner membrane. The outer shell is further divided into three layers and has openings, called pores, extending between the inside and outside of the shell. The highest number of the pores is found around the middle circumference of the egg. The outermost layer is a thin characterless layer made up of calcium and phosphorus, and the middle layer consists of interlocking wedge-shaped calcite crystals with traces of magnesium, sodium, sulphur, and phosphorus. These crystals end in cone shaped knobs on the inner aspect of the shell and then also forms the third inner layer of the shell. The amount of these knobs, called mammillae, is highest at the equator of the egg and becomes lower towards the egg poles. The shell thickness follows the same pattern, with the thickest area located at the equator. The second layer of the egg, the membrane is found right beneath the shell and is embedded in the mammillae. The membrane is composed of numerous fibres, roughly arranged in parallel sheets of varying directions. Each individual

fibre is made up of smaller fibrils clustered together, and forming open channels that run lengthwise through the fibre.

THEMED ISSUE ARTICLE

Addressing multiscale microbial challenges using the Mesolens

Liam M. Rooney, Beatrice Bottura, Katherine Baxter, William B. Amos, Paul A. Hoskisson, Gail McConnell

We provide a brief review of the development and application of the Mesolens and its impact on microbiology. Microbial specimens such as infected tissue samples, colonies surfaces, and biofilms are routinely collected at the mesoscale. This means that they are relatively large, multimillimetre-sized samples which contain microscopic detail that must be observed to answer important questions across various sectors. The Mesolens presents the ideal imaging method to study these specimens as no other optical microscope can, thanks to its unique combination of low magnification and high numerical aperture providing large field-of-view, high-resolution imaging. We demonstrate the current applications of the Mesolens to microbial imaging and go on to outline the huge potential of the Mesolens to impact other key areas of microbiology.

ORIGINAL ARTICLE

Facet type determination based on combined atomic force microscopy and electron backscatter diffraction

Ralf Brüning, Mehrad Hajati, Peter G. Lelièvre, Tobias Bernhard, Sascha Dieter, Grégoire Dietrich

Crystals are usually recognised by having distinct facets at their surface.The study of crystals and facets has a long tradition. To identify a crystal facet, one must know the orientation of the surface relative to

stacking pattern of the atoms within the crystal. Both these tasks can be accomplished separately, but it is difficult to put the two pieces of information together for hundreds or thousands of crystals at a time.

As the dimensions of the electronic circuits become smaller, for example in mobile phones, it is becoming increasingly important to know and control the facets present at the surface of the electrical connections. We measure the topography of the sample surface with Atomic Force Microscopy (AFM) and identify flat parts of the surface as facets. On the same sample, we use a scanning electron microscope with a specialised module (Electron backscatter diffraction, EBSD) to find the directions in which the atoms are stacked below these facets. Combining the data from these two instruments allows us to find the facet types. The mathematical steps needed to do this are described and illustrated. This new technique will allow improved process design and process control in electronics manufacturing and other areas.

REVIEW ARTICLE

Expansion microscopy: A revolution in diagnostic pathology

Snehashish Ghosh, Bhawana

Subedi Sapkota, Roopa S Rao, Shankargouda Patil, Chandini Rajkumar, Surendra

Lakshminarayan

Expansion microscopy (ExM) is a recent discovery in diagnostic pathology and microbiology that promotes a physical magnification of the tissue specimen instead of optical magnification. It not only improves the resolution of the specimen but also enhances the diagnostic precision, and permits nanoscale imaging of the specimen.

Optical microscopy is routinely used in histopathology and microbiology for ages. Due to its simplicity, compatibility with different types of specimens, and ease of operation it is accepted by pathologists. ExM and

its variants have been widely tested in different types of tissue specimens and microbiological specimens and yielded brilliant results.

INVITED REVIEW

Analysis of 3D elemental distribution in nanomaterials: Towards higher throughput and dose efficiency

Alexander Skorikov, Kees Joost Batenburg, Sara Bals

Many advanced nanomaterials rely on carefully designed morphology and elemental distribution to achieve their functionalities. Among the few experimental techniques that can directly visualise the 3D elemental distribution on the nanoscale are approaches based on electron tomography in combination with energy-dispersive X-ray spectroscopy (EDXS) and electron energy loss spectroscopy (EELS). Unfortunately, these highly informative methods are severely limited by the fundamentally low signal-to-noise ratio, which makes long

experimental times and high electron irradiation doses necessary to obtain reliable 3D reconstructions. Addressing these limitations has been the major research question for the development of these techniques in recent years. This short review outlines the latest progress on the methods to reduce experimental time and electron irradiation dose requirements for 3D elemental distribution analysis and gives an outlook on the development of this field in the near future.

THEMED ISSUE ARTICLE

Overcoming the challenges of preserving lipid-rich Cannabis sativa L. glandular trichomes for transmission electron microscopy

Samuel J. Livingston, Eva Yi Chou, Teagen

D. Quilichini, Jonathan E. Page, A. Lacey Samuels

Cannabis glandular trichomes produce and store an

abundance of lipidic specialised metabolites (e.g. cannabinoids and terpenes) that are consumed by humans for medicinal and recreational purposes. Due to a lack of genetic resources and inherent autofluorescence of cannabis glandular trichomes, our knowledge of cannabinoid trafficking and secretion is limited to transmission electron microscopy (TEM). Advances in cryofixation methods have resulted in ultrastructural observations closer to the ‘natural state’ of the living cell, and recent reports of cryofixed cannabis trichome ultrastructure challenge the long-standing model of cannabinoid trafficking proposed by ultrastructural reports using chemically fixed samples. Here, we compare the ultrastructural morphology of cannabis glandular trichomes preserved using conventional chemical fixation and ultrarapid cryofixation. We show that chemical fixation results in amorphous metabolite inclusions surrounding the organelles of glandular trichomes that were not present in cryofixed samples. Vacuolar morphology in cryofixed samples exhibited homogenous electron density, while chemically fixed samples contained a flocculent electron dense periphery and electron lucent lumen. In contrast to the apparent advantages of cryopreservation, fine details of cell wall fibre orientation could be observed in chemically fixed glandular trichomes that were not seen in cryofixed samples. Our data suggest that chemical fixation results in intracellular artefacts that impact the interpretation of lipid production and trafficking, while enabling greater detail of extracellular polysaccharide organisation.

ORIGINAL ARTICLE

Analysing the effect of sodium bicarbonate and glycine air polishing on tooth surfaces with two different imaging methods

V. Merve Balta-Uysal, Kaan Orhan, Ece Irem Oguz, Esra Guzeldemir-Akcakanat

The aim of this study was to compare two different imaging methods by assessing changes caused by sodium bicarbonate and glycine air polishing on the tooth surfaces. Fourteen single root teeth with exposed root surfaces were included into the study. The teeth were randomly divided into two groups: sodium bicarbonate and glycine group. Samples were scanned in a micro-computed tomography (micro-CT) and CAD/CAM (computer-aided design/ computer-aided manufacturing) at baseline and then after air-polishing powder applications, the defect volume values were evaluated. There was a statistically significant difference between mean defect volume values that occurred after glycine and sodium bicarbonate air polishing evaluated with micro-CT and CAD/CAM (p < 0.05). After sodium bicarbonate air polishing, defect volume on enamel surface at maximum power and defect volume on the exposed root surface at medium power values calculated with CAD/CAM were higher. After glycine air polishing, defect volume values on both surfaces at medium power setting calculated with CAD/CAM were lower. Defect volume values on enamel surface at maximum power setting calculated with CAD/ CAM were higher than calculated with micro-CT. We concluded that CAD/CAM cannot provide as accurate results as micro-CT. Glycine-based powder is less abrasive than sodium bicarbonate, especially on enamel surface.

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Journal of Microscopy launching call for papers for new special issues in 2023

We would like to invite attendees of the Microscopy of Semi-Conducting Materials meeting, which is being held 3-6 April in Cambridge, to submit a paper for a special issue of the Journal of Microscopy due to be published in late 2023.

The guest editors of the issue are Thomas Walther (University of Sheffield) and Rachel A Oliver (University of Cambridge).

The deadline for papers is 31 May 2023.

We have also launched a special issue from the new RMS section, Data Analysis in Imaging (DAIM), “Data Analysis in Imaging.”

There is an open call to all the bioimage analysts and data scientists to contribute to the Journal’s special issue with focus on tools and workflows for image analysis and data management in imaging.

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The Journal of Microscopy welcomes papers for two new special issues planned for 2023.

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.

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Member Profiles

Name Gaurav Chand

Tell Us About You?

Gaurav Chand is a second-year PhD researcher in the department of Mechanical, Aerospace and Civil Engineering at the University of Manchester. He has five years of research experience in structural engineering. His research area focuses on developing sustainable concrete (low-carbon concrete) from waste materials.

Why did you become a member of the RMS?

RMS is a prestigious, world-renowned society that provides comprehensive information and updates regarding microscopical tools, which are vital for my microstructural analysis of concrete.

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Membership in RMS provides vast exposure to the international community of distinguished researchers from different fields. It keeps updated with the latest developments in microscopy such as advances in microscopy techniques or novel application areas.

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Nino Pochkhidze

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MS, PhD student of Cellular Neuroscience at Ilia State University and Researcher of Department of Brain Ultrastructure and Nanoarchitecture at Ivane Beritashvili Center of

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Aarti Sagar Patankar

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I am currently pursuing a Bachelor’s degree in Microbiology. I have a keen interest in Genetics and I would like to study CRISPR Cas9 system and its applications. I am an enthusiastic individual, passionate about improving my understanding of the subject. I’m looking for opportunities in the field of Microbiology and Genetics and hoping to contribute to the Society by doing extensive research.

Experimental Biomedicine. Nino’s main interests are behavioral (social activities, cognition, emotional sphere, Microscopes), morphological, ultrastructural and nanoarchitectural effects of inhalants with addictive properties in the organisms of different ages.

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Becoming a member of the RMS will give me insights and keep me updated with ongoing research and developments. I am very excited to be a part of this esteemed society.

How do you feel being an RMS member benefits you?

The RMS will help me to grow in academia as a researcher and broaden my perspective. It will open the doors to new opportunities.

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The RMS Digital Calendar for 2023 got off to a great start in January and February with a pair of fabulous images depicting the beauty of the living world.

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All the images used were submitted to our annual calendar image competition. Congratulations to the winners!

Pine stem

Transverse section of a Pine stem showing resin duct. Equipment Used: Brightfield image using a 5megapixel digital eyepiece camera on a Wild M20 Research microscope

Michael R. Gibson, Northamptonshire Natural History Society & Royal Microscopical Society.

Image Credit: Prepared Biosil slide by the late John Wells

Fungi in spider web

Colourised SEM image of a fungi colony found in a spider web. A special collection method was applied to preserve the tension and the shape of a spider web as well as to allow for sample manipulation and imaging on both sides. The sample was coated with 7nm of platinum on both sides. Image processed and colourised with use of Mountains Map software.

Equipment Used: Quorum Q150V S Plus coater, Tescan Amber FIB-SEM.

Anna Walkiewicz, Quorum Technologies

Investigating the liquid Characterising the surface quality of 3D printed

optical elements using interference reflection microscopy

Student: Ben Watson, BSc (Hons) Physics, 2nd Year

Location: Department of Physics, SUPA, University of Strathclyde, Glasgow, UK

Optical microscopy has used the same manufacturing methods for milling and sanding glass elements to produce high-quality optics for over a century. Recent developments in 3D printing have opened the possibility of printing optical components, but the surface quality of these 3D prints remains unknown. To address this, I designed and printed several planoconvex lenses using a consumer-grade 3D printer, and then experimented with various post-processing methods to improve the quality of the lens surface. I then used a confocal microscope set up in reflection mode to image the topology of the lens surface following different post-processing methods. This method produced constructive and destructive interference patterns on the lens, displayed as fringes corresponding to the axial position of the lens surface. I wrote my own code to reconstruct a 3D visualisation of the lenses using the interference fringe data and then calculated the radius of curvature of the 3D printed lens. This data was compared to the specifications of the CAD model that was printed. By the end of my studentship, I had developed a workflow for manufacturing 3D printed lenses and written a custom analysis script that was used to characterise and quantify their surface quality.

What was the aim of your project?

The aim of this project was to develop a workflow to produce high-quality 3D printed optical elements and to develop a microscopy-based method to characterise the 3D printed lenses.

How did you address the aim?

I began the project using a 3D printer that used digital light processing (DLP) technology to produce 3D printed lenses. The DLP method uses ultraviolet (UV) light to project, layer-by-layer, a slice of the computational model of my lens onto an LCD screen in contact with a resin bath. The UV light caused photo-induced polymerisation of the liquid resin into a solid printed layer matching the projection. The lenses created by this method were initially opaque, which was caused by incident light being dispersed due to the uneven surface of the raw printed layers. The surface quality was improved using different post-processing methods:

• Spin-coating: when a small volume of resin was applied to the apex of the lens and then rotated at high speed to pull the resin evenly over the surface.

• Drop casting: similar to spin coating in that resin was applied to the apex of the lens but, instead of spinning, the lens was left flat and gravity distributed the resin over the surface.

• Hand sanding and polishing: the use of very fine grit sandpaper and then applying clear acrylic polish to progressively remove/smooth surface features.

The lenses produced by each of these methods gave various levels of success. However, a quantitative method to characterise the surface quality of the lens was required.Therefore, the lenses were imaged using a confocal microscope setup for interference reflection microscopy (IRM), where a beam-splitting mirror is placed in the usual position of a dichroic mirror. This method resulted in interference patterns which mapped the surface of the lens, like

Figure 1. An overview of lenses printed during the project. Multiple lenses were printed and processed using different spin coating conditions, tested by IRM, and their surface profile characterised and compared to a glass lens of the same prescription. Each of the 3D printed lenses were printed with a thin raft surrounding the lens to ease handling, these rafts did not alter the optical parameters of the lenses.

that of contour lines on an Ordinance Survey map. The height of the lens surface where constructive interference occurs was only dependent on the order of constructive interference, the refractive index of the medium the lens was in, and the wavelength of incident light. A 3D reconstruction of the lens was created using IRM and compared to their predicted geometry. This data was used to calculate the radius of curvature of the printed lens and compared to the theoretical lens curvature, or compared to a precisely manufactured glass lens of the same design specifications.

What did you find out?

The surface profile of the lenses was measured using the interference fringes present in the IRM data, which corresponded to the axial position of the curved surface. The axial position at which constructive interference occurred was used to create a 3D reconstruction of the lens using a dedicated python script. The curvature of the lens was then calculated and compared to the

theoretical values.

During this project, most of my time was spent on the spin coating method. I found that it was possible to manufacture lenses with high surface quality, but the results could be inconsistent depending on the volume of resin, spin-time, spin-speed, and the method of resin application.

Hand polishing seemed promising at first, as this is the routine method for professional manufacturers to process glass lenses. However, these lenses were impossible to analyse as there was no interference pattern due to irremovable resin surface roughness, if lenses were instead polished by machinery, it may have yielded a better result.

The last method that I investigated was drop casting which seemed like a promising way to create a lens, however, needs to be investigated further to test the reproducibility of the method.

The model lens that we used was the LA1131 from Thorlabs which had a radius of curvature of 25.8

Figure 2. Characterisation of the surface quality and calculation of curvature from IRM data. (a) A commercially sourced glass lens with a known radius of curvature of 25.8 mm. The IRM image (left) shows the concentric rings of constructive and destructive interference corresponding to the topography of the lens surface. A 3D reconstruction is presented (middle) following processing of the IRM data (black) and compared to the theoretical curvature (green). The radius of curvature (R1) and surface quality were measured by calculating R1 in a radial sweep and presented as a histogram (right), with the theoretical radius of curvature denoted by a green dotted line. The spread of the data gives an indication to the surface quality of the lens. (b) the same as above, but for a 3D printed lens of the same prescription.

mm. The spin coating method produced the best quality lenses, with radii ranging from 27.7 mm to 32.1 mm, slightly larger than the manufacturer’s values. The lenses that I drop cast had radii of curvature of 22.8 mm and 29.5 mm. As mentioned, the polished lenses were impossible to analyse with this method.

What did you learn from participating in this project?

Over my time working on my studentship, I learned a lot of practical skills such as how to operate a 3D printer and how to acquire IRM images using a confocal microscope. I was also able to gain an understanding of how to operate advanced imaging equipment, and expanded my understanding of the theory behind microscopes as well. I was able to apply and develop my coding skills when analysing the images taken using the microscope, which created a new method for characterising the surface of 3D printed lenses.

I gained experience working in a wet lab as part of a multidisciplinary research team which I would not have had the opportunity to do outside of this studentship. Also, at the end of the studentship I had the opportunity to present my data at a research seminar, which taught me how to create a presentation targeted toward an interdisciplinary audience.

How has this project affected your long-term goals?

Doing this project has given me a window into what it’s like to work in research as part of the academic community, something which I was not able to get from studying for my degree alone. This project has expanded my network and put me in contact with academics, which will undoubtedly be useful in my next few years at university. Overall, this project has made me consider the possibility of having a career in academia more seriously.

Ben Watson

Investigating the liquid phase separation of Rubisco and linker proteins in vitro

Student: Catherine Read

Supervisor: Alex Payne-Dwyer

Location: School of Physics, Engineering and Technology, University of York

Most of the oxygen we breathe finds its origins in marine algal photosynthesis. These algae organise their Rubisco, the enzyme responsible for capturing carbon dioxide and releasing oxygen, into a liquid droplet called the ‘pyrenoid’. The algae then concentrate carbon dioxide into the liquid, and the reactions can run faster. The pyrenoid improves the efficiency of the oxygenproducing reaction by around 60% compared to the same reaction in plant leaves. Understanding the phase-separation process will help scientists introduce these carbon concentration mechanisms into plants, with the hope of increasing crop yields to help with food security. However, for these liquid droplets to form at all, Rubisco requires a counterpart ‘linker’ protein. We don’t know what minimal properties of a linker protein are needed for pyrenoids to function this way. And why a liquid? What are the resulting properties of the droplet that drive this incredible boost in photosynthetic output?

I created pyrenoid-like droplets by mixing these proteins outside the confines of a cell. I used a specialised microscope to measure the droplet refractive index (the amount that light is slowed down in a material) as a starting point for estimating the amounts of Rubisco and linker concentrated inside the droplets.

What was the aim of your project?

Liquid phase separation is a fascinating strategy that cells borrow from physics to speed up vital reactions. I aimed to take CO2-fixing enzyme and linker from Chlorella algae, to mix them to get droplets, and to correlate the droplet size, fluorescence (labelled

protein) and refractive index (total protein) with composition.

How did you address the aim?

The project started with micromolar amounts of purified Rubisco and linker proteins. I mixed them

at specific ratios. After confirming the presence of droplets in brightfield, I then put the droplet mixtures in a Tomocube microscope, which performs label-free holotomographic reconstruction of the sample volume. I converted the image of optical phase shift to an estimated map of the refractive index of the droplets, using a calibration with two well-defined samples: first, the neat buffer without protein, and second, polystyrene microbeads in glycerol. While the peaks in refractive index were distinct from the background, artifacts of the reconstruction were problematic as some droplets were very small. With high-precision coverslips, these artifacts became more manageable. However, I still needed a way to confirm which signals were

droplets. I doped the mixtures with 5% or 100% GFP-tagged linker protein so that I could observe the droplets under epifluorescence and compare with the maps of refractive index. I wrote macros in FIJI to determine fluorescence thresholds that correctly segmented droplets from surrounding buffer. This eliminated remaining artifacts of the tomographic reconstruction. I could then map the refractive index inside the droplets, which I then used as a proxy for the local protein concentration.

What did you find out?

Although the fluorescent signal was weak, the relative lack of tagged linker outside the droplets meant a very low fluorescent background as well. I

Figure 1. Liquid droplets of protein show inclusions of the dilute buffer phase when agitated. Widefield epifluorescence image (green) and refractive index map (magenta) and merged overlay (left). Registered line profiles show good correlation (right). Scale bar: 5 μm.

was able to segment each droplet in the fluorescence channel and estimate its projected size. After I registered the fluorescent and refractive index images, I was able to confirm that they matched up with good overlap (Figure 1, lower left). I could then identify which areas of the images corresponded to the same droplet and look at correlated data across droplet populations.

A key question was whether droplets produced with 100% GFP-tagged linker had the same properties, or if the fluorescent tag somehow perturbed the phase separation process. I observed that droplets formed using 5% GFP-tagged linker (Figure 2, left) were many times smaller than those with 100% GFP-tagged linker (Figure 2, right). This suggests that tags encourage phase separation, probably by increasing the attractive interactions of the linker with Rubisco. When agitating the 100% tagged mixture with a pipette, the regular droplets formed beautiful double emulsions (Figure 1). Hopefully, the reluctance of these droplets to fuse might tell us something about their structure or surface properties in future work.

No strong correlation was found between droplet size and refractive index, at least for very small droplets with 5% GFP-tagged linker (Figure 2, left). However, using 100% GFP-tagged linker to generate a larger range of sizes (Figure 2, right), I found the

average refractive index rapidly increased to a high ‘plateau’ for droplets larger than a micron. This might be explained by a thin surface layer that is lower in refractive index than the droplet core, though this isn’t clear from the line profiles (Figure 1, lower right). Inside the pyrenoid, the Rubisco enzyme is expected from reaction models to a hundredfold more concentrated than in the buffer. In agreement with this, I measure a refractive index range that corresponds to a Rubisco enrichment of between 100- to 400-fold inside the droplets.

What did you learn from participating in this project?

During my project I have received training on a number of different microscopes as well as the Tomocube, including Slimfield (single molecule tracking in the Leake group at York), Zeiss LSM confocal and epifluorescence microscopes. The Tomocube had a difficult learning curve, but I became more proficient towards the end of my project. I was able to acquire some good images and it helped with my general understanding of research microscopy and scientific instruments. I have a background in biochemistry and hadn’t realised that such a large component of microscopy at this level is downstream image analysis, which I

Figure 2. The modal refractive index measured inside each droplet against its cross-section size, using (left) 5% GFP-tagged + 95% native linker, or (right) using 100% GFP-tagged linker.

applied through use of FIJI software to compare images and put numbers to different measures. I have also experienced training in the wet lab such as preparing the protein mixtures, mounting different slides, and preparing bead samples for calibration of different microscopes.

How has this project affected your long-term goals?

While I am undecided what path I want to pursue in the future, as there are so many options, this project has definitely shown me that research is a very rewarding line of work. During research many different challenges are encountered - my progress was quite limited by the short timespan of the project and the purified protein available. Yet time is spent finding different solutions and ways around these obstacles. It is also a very friendly

environment as part of a research team; everyone shares their findings and helps each other out with their different projects, all working to reach one final result. Doing this project has confirmed that I enjoy interdisciplinary science so that is definitely something I would wish to continue in future work.

Contacting the Royal Microscopical Society

Catherine Read

Using image processing to quantify biologically relevant information in fluorescence microscopy images

Student: Hale-Seda Radoykova

Supervisor: Dr Siân Culley

Location: Randall Centre of Cell & Molecular Biophysics, King’s College London

Accurately detecting and measuring structures in fluorescence microscopy images is important yet challenging. One of the interests of the Culley lab is to develop image analysis techniques to help researchers with these tasks. The aim of this summer project was to acquire images of fluorescently labelled tubulin in Schizosaccharomyces pombe cells and to produce an image processing pipeline adapted to quantifying the microtubules in these images. One approach to detecting these linear structures was based on thresholding and skeletonisation, using plugins available in Fiji/ImageJ. This workflow yielded generally satisfactory results in the simplest case but was prone to creating artefacts and to loss of information. Crucially, this method could not cope with intersecting lines. As a more powerful alternative, the linear Hough transform was used to detect straight lines. The implementation in Python was efficient and overcame some of the drawbacks of the previous method. However, it required significant post-processing to faithfully detect multiple lines in images. Unfortunately, it was only feasible to parametrise simpler instances of straight lines, and the method could not be extended to e.g., curved lines. Deep learning approaches could be an exciting front for future work on this problem.

Research question

How can image processing tools be used to quantify the linear structures present in fluorescence

microscopy images of labelled tubulin in Schizosaccharomyces pombe (fission yeast)?

Aim

Develop a coherent image processing pipeline suited to S. pombe data.

How did you address the aim?

Wet lab work

The wet lab component of the project included culturing and maintaining S. pombe strains endogenously expressing sfGFP-Atb2.We were able to acquire some widefield fluorescence microscopy images.

Complementary to this, our collaborators at the Dey lab (EMBL, Heidelberg) kindly provided several stacks of time-series maximum-projected confocal microscopy images with a wide field of view containing numerous S. pombe cells in the exponential growth phase (typically 190-210 cells per image). The bundles of microtubules in these cells looked like lines which could be parametrised.

Computational methods

The first part of the computational method implemented a simple skeletonisation pipeline using ImageJ/Fiji [1]. Manually selected 128x128 ROIs (regions of interest) were copied and sorted into several categories for further processing. ImageJ macro scrips were used to apply the same line detection procedure to each ROI: a smallradius Gaussian filter, auto-thresholding using the “triangle” algorithm, and erosion of the binarised image, followed by skeletonisation. This yielded binary images with a one-pixel wide outline of

the detected structures which could be measured within ImageJ.

One parametrised approach for straight lines is using the linear Hough transform. The transform represents each point in Cartesian coordinate space as pair of parameters in Hough space [2]. Straight lines were parametrised using polar coordinates, as for lines of the form The linear Hough transform was implemented in Python, and output was refined using libraries, such as scikit-image [3]. The algorithm was modified to detect lines based on pixel intensity weights from raw non-binary data.

What did you find out?

Fiji pipeline

The pipeline in Fiji/ImageJ had the advantage of being relatively simple to implement and automate. It worked relatively well for detecting single straight and curved lines with consistent signal. Line endpoints and length were easy to obtain directly from the binary images.

However, with thresholding and skeletonisation techniques, there were multiple ways of obtaining satisfactory results by tuning one or more parameters. Intrinsic image heterogeneity led to loss of information along the image periphery. It could also break up lines with inhomogeneous intensities (Figure 1a, orange).

Skeletonisation was prone to creating artefacts; for example, parallel lines were sometimes blended together (Figure 1b, yellow). One significant

Figure 1. Artefacts resulting from the thresholding and skeletonisation process in two consecutive time frames of the same field of view. (a) Inhomogeneous lines could be interrupted (orange); (b) parallel lines could be blended together (yellow); intersecting lines cannot be skeletonised and labelled faithfully (blue). All scale bars = 2 μm.

disadvantage for downstream processing was the constraint that intersecting lines could not be labelled as separate objects (Figure 1, blue).

Hough transform

The linear Hough transform detected straight lines in unprocessed raw and binarised data alike. Working with raw data had the advantage of not losing information to processing. This workflow was able to detect interesting lines without being sensitive to interrupted lines or lower contrast.

The disadvantages of this method were related to the postprocessing of the detected lines. A local peak detector was used to identify multiple lines in Hough space; this required manual thresholding. In raw images, noise was sometimes picked up as a line by the local maximum detection algorithm, and multiple lines around the same true line were reported (Figure 2a). These had to be curated by fitting Gaussian curves to the line intensity profiles (Figure 2b). Parameters extracted from the Gaussian fits could be used to estimate the positions of line endpoints (Figure 2c).

The Hough transform could be used for simpler cases; however, handling exceptions and artefacts detected by the algorithm quickly became laborious. Extending the linear Hough transform to fit curved lines of a generalised form was not trivial and was not practical to implement here.

Future work

As the parametrised method for microtubule quantification in S. pombe fluorescent microscopy images could only quantify the simplest instances, future work could build upon this by covering the more complex scenarios and work in the full field of view. One intriguing possibility would be building a convolutional neural network (CNN) based on the popular U-net architecture [4]. Exciting previous work on the two-dimensional problem of nucleus segmentation has extended the U-net pixel-to-pixel mapping to pixel-to-shape mapping for instance segmentation based on star-convex objects [5] or splines [6]. Finding a suitable equivalent for predicting lines would be an exciting challenge.

Figure 2. Lines detected by linear Hough transform. (a) Raw image with detected lines and their corresponding peaks in polar parameter space; (b) the intensity profiles of the detected lines; (c) fitted Gaussian curves (in black) and their full width (in grey).

Conclusion

This summer project feeds into the broader aims of the Culley lab to quantify structures in S. pombe and other experimental systems. Segmenting geometrically constrained objects is non-trivial and can have important implications for biological data analysis. For example, having a pipeline which accurately quantifies microtubule number, length, or curvature could enable researchers to make crosscondition comparison of microtubule properties in different experimental settings. This would be impossible or unreliable with a pipeline sensitive to noise or tending to produce artefacts.

What did you learn from participating in this project?

I thoroughly enjoyed working on my summer project at the Culley lab! On the one hand, it helped me consolidate my wet lab skills while also complementing my computational skillset with image processing and fluorescence microscopy image analysis techniques. I familiarised myself with Fiji/ ImageJ macros and gained programming experience in using Python for image processing, analysis, and visualisation. It was also rewarding to persevere through multiple challenges both in the lab, trying to keep the yeast cultures happy, and in debugging my code. Decidedly, the highlight of the entire project for me was the moment in the microscopy room when we finally saw the fluorescently labelled structures moving in our living yeast cells. This has been an unmatched learning experience, and I am very grateful to Dr Culley and the RMS for making it possible!

How has this project affected your long-term goals?

In the immediate future, I am beyond excited to learn and do more fluorescence microscopy for my integrated master’s project next year.The aim will be to use super-resolution microscopy techniques to

give a quantitative comparison of actin remodelling in CAR-T cells vs native T cells. With the skills gained over the summer, I feel much more prepared and inspired for the upcoming year.

Working with Dr Culley has been an absolute pleasure! We are currently looking into short-term grant opportunities, as I would love to return to her group as a research assistant in the near future. Finally, I am grateful for this wonderful opportunity, as not only did I meet inspiring researchers this summer, but I believe that quantitative microscopy might be the field to shape my postgraduate studies and career too.

References

[1] Schneider, C., Rasband, W. and Eliceiri, K. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9, 671–675 (2012). https://doi. org/10.1038/nmeth.2089

[2] Ballard, D.L. Generalizing the Hough transform to detect arbitrary shapes. Pattern

Hale-Seda Radoykova.

Recognition 13:2, 111–122, (1981). https://doi. org/10.1016/0031-3203(81)90009-1

[3] van der Walt, S., Schönberger, J.L, NunezIglesias, J., Boulogne, F., Warner, J.D., Yager, N., Gouillart, E., Yu, T. and the scikit-image contributors. scikit-image: Image processing in Python. PeerJ 2:e453 (2014). https://doi. org/10.7717/peerj.453.

[4] Ronneberger, O., Fischer, P. and Brox, T. U-Net: Convolutional Networks for Biomedical Image Segmentation. Medical Image Computing and Computer-Assisted Intervention –MICCAI, (2015). Lecture Notes in Computer Science, vol 9351. Springer, Cham. https://doi.

org/10.1007/978-3-319-24574-4_28

[5] Schmidt, U.,Weigert, M., Broaddus, C. and Myers G. Cell Detection with Star-Convex Polygons. Medical Image Computing and Computer Assisted Intervention - MICCAI, Granada, Spain, (2018). https://doi.org/10.1007/978-3030-00934-2_30.

[6] Mandal, S. and Uhlmann, V. Splinedist: Automated Cell Segmentation With Spline Curves. 2021 IEEE 18th International Symposium on Biomedical Imaging (ISBI), 1082–1086, (2021). https://doi.org/10.1109/ ISBI48211.2021.9433928

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ANALYZERS FOR ELECTRON MICROSCOPY

The right angle for better analysis

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From the RMS President

Dear Readers,

I would like to wish you all a belated ‘Happy New Year’ – this being our first issue of infocus in 2023. I hope you have had a great start to the year, and that any resolutions you may have made have been firmly kept so far!

It has certainly been a busy couple of months for the RMS following the Christmas break, with many of our popular, annual meetings and courses taking place. We kicked off in January with a well-attended UK Light Microscopy Facility Meeting 2023 in Cambridge, followed by the Flow Cytometry Facilities Meeting 2023 in London. We also linked up once again with colleagues in Australia and Canada for further instalments of the Expansion Microscopy User Group Meeting. This is a great new venture bringing together researchers around the world and enabling the rapid uptake of this technology. The RMS is delighted to provide administrative support for this initiative, including registration and correspondence with attendees.

As I write, we are eagerly anticipating the resumption of the International Microscopy Lecture Series on 15 February, with a talk from Professor Frances Ross, titled Microscopy in motion: understanding how crystals grow through electron microscopy movies We are so pleased to be continuing our partnership with a number of our fellow microscopy societies in delivering these online lectures, and you can find out more about the series, including upcoming lectures, here. The last days of February are also due to feature the ever-popular EM-UK 2023 at London’s Natural History Museum and the return of All Things Cryo in Nottingham - a great course for those keen to explore cryogenic microscopy methods, first held in 2021.

Alongside all this activity, our attentions are now turned firmly towards mmc2023, taking place at Manchester Central once again in early July. It has been four years since we last convened in person for this great event – which feels like an age! The Congress is always eagerly anticipated, but, as they

say, absence makes the heart grow fonder, and I am detecting an extra-special buzz about our international flagship event this year.

Our rich and varied scientific programme at mmc2023 will cover all areas of microscopy, imaging and cytometry – virtually guaranteeing something of interest for all who use some form of microscopy across the sciences. Likewise, the quantity and quality of technology and equipment displayed by our exhibitors never fails to impress. It is worth noting that admission to the exhibition is absolutely free, and well worth the visit to Manchester Central on its own!

If you have not attended an mmc before – or even if you attend regularly - I would strongly urge you to take a look at everything that is on offer this year by visiting the official website: mmc-series.org.uk.You can also read more in this issue on p50.

Also in this issue, we present reports from each of our 2022 Summer Studentship recipients –undergraduates who completed microscopy projects last summer with financial support of up to £2,000 each from the RMS. It is always a pleasure to read about the work they have performed, and to see the scientific contributions already being made by microscopists at the very beginning of their careers.

Finally, I would like to congratulate Professor J Paul Robinson – or ‘JPR’, as he is known to many - on becoming the latest RMS Honorary Fellow. It is a richly deserved accolade for a brilliant scientist who has made worldwide contributions to the field of Cytometry.

So here’s to the rest of 2023! I look forward to meeting many colleagues and friends through our shared passion for microscopy in the coming months, and I encourage you to get involved in Committees and Sections in our great Society – the oldest microscopy society in the world!

Professor Grace Burke, RMS President

PhD student Nadine revamps web resources during internship

The RMS would like to say a big ‘thank you’ to Nadine Field, who has been carrying out some improvements to the Society’s website as part of a recent internship.

Nadine, who is currently studying for a PhD in plant biology at Oxford Brookes University, began a parttime work placement at the Society in September last year. Since then, she has repackaged a huge number of online microscopy resources – making them more user-friendly and accessible to RMS members and the wider public.

She is one of the first PhD students to receive financial support from the Chris Hawes Memorial Fund, established at Brookes following the former RMS President’s death in 2019.

Nadine said: “As part of my PhD course we have to undertake an internship, and I knew that Chris was a really well-known and respected figure within the Society during his life. My supervisor thought it would be really nice if I did my internship with the RMS, to carry on that link.”

Nadine’s work has involved curating and reconfiguring a large number of online training resources and links to software packages - much of which was compiled during the Covid pandemic. While previously these existed as a list of links, they are now available as a user-friendly resource library. She has also reorganised the online infocus ‘backfile’ of articles – stretching back to 2006, when the first issue of the magazine was published. The articles can now be accessed much more easily, with readers able to use search terms to identify the content most likely to interest them.

Nadine said: “As a PhD student, I have to do a lot of very focused reading in my subject area, which can be quite limiting when you are trying to find more out-of-the-box ideas. So going through all the resources and articles meant I got to read many articles from outside areas, which was super-fun

because I don’t get to do that much. I found a lot of image analysis and online resources that I wouldn’t have known about, which has been really interesting and quite beneficial for my PhD."

Despite coming to the end of her internship, Nadine hopes to continue her association with the RMS – and has already volunteered to help out at the Botanical Microscopy Meeting 2023 in Norwich in April.

“I will get to learn about conference organisation and industry, as well as present some of my work”, said Nadine. “It will be my first ever conference which is super-exciting. Everyone at the RMS is so nice and has made me feel so welcome. I’ve really enjoyed my time here and would love to carry on volunteering at conferences in future if the RMS will have me! Hopefully, when I’m a bit further into my PhD I can also send in some interesting articles for infocus.”

Nadine Field

Professor J Paul Robinson announced as new RMS Honorary Fellow

‘JPR’ to receive Society’s most prestigious accolade for contributions to cytometry

The RMS is delighted to announce Professor J Paul Robinson as its latest Honorary Fellow.

Joseph Paul Robinson, known as Paul and even ‘JPR’, has made a huge contribution worldwide to the field of Cytometry. This includes his list-serv ‘Purdue Cytometry List’ which is utilised by over 4,500 cytometrists, his work to teach Cytometry in Africa through his Cytometry for Life organisation, and his patented Cytometry technologies.

JPR is the Distinguished Professor of Cytometry at Purdue University. Indiana, USA. He is the past president of the International Society for the Advancement of Cytometry (ISAC) (2006-2008), is Editor-in-chief of Current Protocols in Cytometry, Associate Editor of Histochemica et Cytobiologica and past Associate Editor of Cytometry Part A. He has published over 200 peer-reviewed publications, 36 book chapters, edited 10 books and given hundreds of international lectures and conference

presentations. JPR contributed a keynote speaker at the flowcytometryUK meetings in 2007 and 2016.

JPR has been active at the forefront of flow cytometry and microscopy development for several decades. His most notable achievements have been in developing innovative technologies such as spectral cytometry using multiarray PMTs (which has been commercialised by Sony and Thermo Fisher); developing optical tools for quantitative fluorescence measurement; advanced classification approaches for clinical diagnostics and bacterial classification; high content, high throughput screening technologies - specifically novel analytical software; and most recently, the development of single photon detectors in flow cytometry which would enhance the limit of detection.

JPR was the founder of ‘Cytometry for life’, a notfor-profit charity launched to focus attention on the need for low-cost CD4 technology for developing countries (www.cytometryforlife.org). To further draw attention to the issue of low-cost CD4, he successfully summited Mount Everest in 2009.

JPR’s unique contributions to Cytometry over the past 25+ years have been global. He has been recognised internationally for his contributions, including being awarded the ISAC Membership award in 2014 and the Distinguished Service Award in 2019.

Support the RMS by leaving a legacy in your will

Help bring microscopy to children and support those starting out in the field

As a Charity, donations and gifts help the RMS to continue its outreach work, bringing microscopy to children and helping those just starting out in the field to get the springboard they need to progress successfully.

For example, your donation could:

• support our Microscope Activity Kit scheme for primary schools, introducing young

children to microscopy for the first time

• support our Summer Studentship scheme, in which undergraduates complete a summer microscopy project in their area of interest

• provide Travel Bursaries for students and early career researchers who have been accepted to present their work at a conference but do not have the funds to get there

Find out more and download our Legacy leaflet

Changes to wording of RMS ByLaws approved

New wording adopted to promote inclusivity and diversity

A number of changes to the wording of the current RMS By-Laws have now been approved by the Privy Council – the formal advisory body to His Majesty the King.

This follows proposals put forward at the most recent RMS Annual General Meeting in September 2022, and subsequently approved by the RMS Board of Trustees.

The changes have been made to reflect the RMS’s ongoing efforts to become a more accessible and inclusive society, and to encourage diversity in all its activities.

The RMS Charter, By-Laws and Rules documents are available to view on the RMS website.

You can also read more about the Society’s Equity, Diversity, Inclusion & Accessibility (EDI&A) Group

Molly Hair wins 2022 RMS Early Career Speakers’ Competition!

Second-year PhD student wins judges’ vote during SEMT One Day Meeting 2002

Oxford Brookes Centre for Bioimaging, was among several Early Career microscopists who gave an oral presentation at December’s SEMT (Society of Electron Microscopy Technology) One Day Meeting.

She won over the judges with her captivating talk about her work on whole cell segmentation Volume EM, which is providing quantitative information about various organelles within the single-celled parasite Leishmania. The full title of her presentation was ‘Patterns of organelle duplication and segregation in the Leishmania mexicana cell cycle revealed by 3D electron microscopy segmentation of mitochondrial cristae in volume electron microscopy data’.

Our warmest congratulations go to PhD student Molly Hair, winner of the 2022 RMS Early Career Speakers’ Competition.

Molly, who is in the second year of her studies at the

The RMS Early Career Speakers’ competition is always a highlight of the SEMT meeting. The aim is to encourage those relatively new to microscopy to present their work in front of a scientific audience – delivering a short and well-thought-out talk in an economical style.

Imaging Cell Dynamics

14-17 May 2023 – Pestana Palace Hotel, Lisbon, Portugal

Register now

Abstract submission deadline: 3 March 2023

Final registration deadline: 31 March 2023

biologists.com/celldynamics2023

#celldyn2023

Organisers

Erika Holzbaur

Jennifer Lippincott-Schwartz

Rob Parton

Michael Way

Speakers

Tal Arnon

Francesca Bottanelli

Lucy Collinson

Gaudenz Danuser

Meng Meng Fu

Ricardo Henriques

Wanda Kukulski

Melike Lakadamyali

Christophe Leterrier

Suliana Manley

Caren Norden

Verena Ruprecht

Yannik Schwab

Erdinc Sezgin

Daria Siekhaus

Yvette Wong

Organised by

Image: “The Vimentin Cytoskeleton” Andrew Moore, Howard Hughes Medical Institute, USA

Destination Liverpool: The European Light Microscopy Initiative is returning to the UK in 2024

An organising team of Early Career Researchers (ECRs) have been tasked with organising one of the largest light microscopy meetings in Europe after successfully bidding for the UK to host elmi2024. The ECRs will be supported and mentored by seasoned organisers to pass the gauntlet onto the next generation of imaging scientists and deliver this much-loved, high-impact meeting.

The 2022 European Light Microscopy Initiative (elmi) meeting was held in Turku, Finland, from 0712 June 2022. Since then, we’ve read reports on the success of the meeting from Beatrice Bottura (University of Strathclyde, UK) and Ciarán ButlerHallissey (Aix-Marseille Universite, France) in the September 2022 issue of infocus. The meeting was a great showcase of the latest in optical imaging by all accounts.

A recurring feature of the elmi programme is the opportunity for delegations of prospective organisers to bid for a future elmi meeting to be hosted in their city. Normally these delegations are comprised of established academics; leaders in their respective areas of optical imaging poised to ensure that the next elmi is as good as the last. Prior to elmi2022, we received a proposal to shake things up a bit.

A group of Early Career Researchers were approached to consider submitting a bid to host

elmi in the UK in 2024. This group includes Siân Culley (King’s College London, UK), Joëlle Goulding (University of Nottingham, UK), Jessica Valli (HeriotWatt University/ESRIC, UK), and me (University of Strathclyde, UK). The USP of our bid was that the gauntlet would be passed to the next generation of researchers to take care of organising, but crucially we would be supported by buddying up with a team of established researchers with lots of experience. Our mentor organisers are Gail McConnell (University of Strathclyde, UK), Peter O’Toole (University of York, UK), Tim Self (University of Nottingham, UK), Marco Marcello (University of Liverpool, UK), and Kurt Andersen (Francis Crick Institute, UK).

Those above in attendance at elmi2022 presented the UK bid; Gail and I took to the stage to present a vision for elmi2024 in the UK. We proposed Liverpool as the host city, with its excellent transport links from multiple international airports

and cross-country rail links. The cultural draw of Liverpool was also a factor we leaned heavily on. The obvious links are there, with football, racing, The Beatles, and the arts; but there’s also Liverpool’s rich maritime history and engineering past.

After some final refinements to our bid, we were successful – elmi2024 will be hosted in Liverpool’s Arena and Convention Centre (ACC). This will be a four-day meeting with a heavy focus on research from Early Career scientists, held from 3 – 6 June 2024. The ACC was selected owing to its excellent accessibility, central dockside location, sprawling 1700m2 exhibition space, and over 15 available workshop rooms. An elmi staple, we haven’t forgotten the traditional Academia Vs Industry football match, and we have also proposed a meeting banquet at The Rum Warehouse… which has plenty of space to accommodate the postbanquet dancefloor and DJ.

Of course, none of this would be feasible without

the backing of the RMS. The RMS agreed to support us as our Professional Conference Organiser, scouting out potential conference venues, ensuring accessible routes to allow industry vendors to bring their latest instrumentation to the tradeshow, and outlining and underpinning the finances for a UK elmi. The RMS have a demonstrable track record of successfully organising elmi, with the organisation of five previous elmis being supported by them –York (2007), Glasgow (2009), Leuven (2012), Dublin (2018), and an online elmi (2021).

We hope to see you in Liverpool for elmi2024, but until then, you can keep up with the key dates and developments via the RMS Events Calendar.

Liam Rooney Postdoctoral Researcher, University of Strathclyde Chair of the RMS Early Career Committee

Turku, Finland was the location for elmi2022

The Microscience Microscopy Congress (mmc) is back with a bang in 2023! One of the biggest events of its kind in Europe, mmc2023 (incorporating EMAG 2023) will bring you the very best in microscopy, imaging and cytometry from across the globe. With a huge conference, a world-class exhibition, workshops, social networking opportunities, an international Imaging Competition and more, it’s simply the place to be for anyone who uses a microscope for work, study or pleasure.

Back at the superb Manchester Central conference centre for the first time since 2019, the flagship event of the RMS will be taking place fully in person, providing the perfect opportunity for the scientific community to come together, make connections and share their research. As always, many of the leading companies in microscopy and imaging will be on hand to demonstrate the very latest equipment and technology at the exhibition.

RMS President Professor Grace Burke said: “We are thrilled to be back in Manchester for a fully in-person mmc2023. Nothing beats the buzz of a busy conference hall and exhibition, and the opportunity to catch up with colleagues and friends from right across the global microscopy community.

“With preparations now well advanced, the excitement is certainly building – all the more so because of the four-year gap since we last convened at Manchester Central.We are confident that this year’s event will be one of the best ever microscopy conferences, with a superb scientific programme, world-class exhibition and a host of other great features.”

Registration is now open, and all the latest information can be found on the official mmc-series website: https://www.mmc-series.org.uk

Conference sessions –something for everyone

mmc2023 (incorporating EMAG 2023) will feature no fewer than 36 conference sessions across six parallel streams – with an incredible cast of speakers and supporting poster sessions. The blockbuster programme will cover the full range of latest techniques, applications and hottest emerging topics in microscopy, imaging and cytometry.

The conference programme will also incorporate EMAG 2023, organised by the Institute of Physics’s Electron Microscopy and Analysis Group (EMAG) –with multiple sessions taking place throughout each of the three conference days.

With a huge wealth of scientific content under one roof, we can promise something for all our delegates.

Check out the provisional Conference Programme

Find out more about our Plenary Speakers

Free Exhibition

Meanwhile more than 60 exhibitors, including some of the biggest names in microscopy and imaging, will be on hand to discuss their latest products and give practical demonstrations – including free company workshops. The event also provides important exposure for a number of smaller companies.

mmc delivers the perfect opportunity for exhibitors to reach out to the global microscopy community and showcase a quality and quantity of equipment that never fails to impress.

Much of the floor space has already been snapped up following a fantastic response from companies when sales opened in late 2022. However, bookings remain open and if you would like more information please contact Dawn Hopkins or Nick Cameron.

Entry to the exhibition is completely free for visitors. Check out the current exhibitor list

Satellite meetings and bonus events

mmc2023 (incorporating EMAG 2023) will bring together a number of smaller meetings, allowing you to meet with colleagues working in your field as well as with cross-disciplinary peers, all at the same event. They include:

Pre-Conference Workshops (including AFM & SPM, EMAG and ImageJ) - Monday 3 July.

BioImagingUK Meeting - Monday 3 July

Join the UK Bioimaging community to discuss priorities and strategies in national infrastructure, technology development, training, careers and ways to share knowledge across different disciplines.

(Free to attend)

Early Career Symposium - Monday 3 July

A great opportunity for students, postdocs and early career professionals to network and showcase their research ahead of the main mmc2023 conference

Super-Resolution Workshop - Friday 7 July.

Scientific Imaging Competition

The long-established RMS International Scientific Imaging Competition is winging its way once again to mmc. An eye-catching gallery of the shortlisted images – covering all the sciences and microscopical techniques - will be on display throughout the event, with the winners announced during the conference. The competition is always a highlight of the Congress, bringing an ‘art-meets-science’ flavour to proceedings.

Learning Zone

The RMS will be bringing its ever-popular Learning Zone to mmc, with experts on hand to share their knowledge with visitors and provide demonstrations.

RMS staff and volunteers will also be available to speak to visitors about the work of the Society and the benefits of membership.

Among the attractions at this year’s Learning Zone will be a set of beautiful antique slides donated to the RMS – and discussed by Dr John Hutchison Hon FRMS in this issue on page **.

Register now!

Registration is now open, and all the information on rates, accommodation and transport can be found on the official mmc-series website: https://www. mmc-series.org.uk/

We look forward to seeing you there!

Submit your image to the RMS Scientific Imaging Competition!

Do you have an image (or short video) you would like to share with the microscopy community at mmc2023?

The RMS Scientific Imaging Competition welcomes both images and videos from all microscopy disciplines. Shortlisted entries across seven different categories will be put on display at mmc2023, where the winners will also be announced.

It’s the perfect opportunity for microscopists from across the world to showcase their technical and artistic skills, while revealing the beauty of the microscopical world.

Submissions are now open! The deadline for competition entries is 2 May 2023.

A selection of winning entries from the 2021 Scientific Imaging Competition. View the full gallery and find out more about these images

The Expansion Microscopy User Group: Cultivating a new scientific community

Having first been put forward as a sample preparation tool in 2015, expansion microscopy is something of a ‘new kid on the block’ in scientific research. The process, involving the physical enlargement of a biological sample by the introduction of a polymer network - thereby enabling the analysis of small structures with standard microscopes – has numerous applications in medical research and other fields. Aware of its enormous potential, scientists across the world are now experimenting with the technique. But how can this growing body of knowledge be harnessed effectively?, and how to spread best practice - alerting budding expansion microscopists to the many pitfalls of the preparation process along the way?

Over the last year, discussions within the light microscopy community in North America have given rise to the Expansion Microscopy User Group – organised by Canada BioImaging (CBI) and BioImaging North America (BINA), and now including a counterpart initiative based in Australia (in conjunction with the University of Melbourne). The initiative receives administrative support from the RMS – overseeing the registration process for meetings and correspondence with attendees.

Virtual meetings are hosted by Canada and Australia to offer workable meeting times for everyone using the emerging technique, right across the globe. The discussions are recorded and uploaded to YouTube, enabling all attendees to gain insights from community members attending both meetings. Alongside this, a growing database of information and online discussion is collated

and made available to members of the group. The group currently consists of 102 members from 23 different countries.

Among the Canada-based organisers is Associate Professor Claire Brown, Director of the Advanced BioImaging Facility (ABIF) at McGill University.

She explains: “We wanted to get into expansion microscopy, and I didn’t have any background in the technique myself. In that situation people will traditionally go to the relevant papers explaining a particular technique, and then troubleshoot in the lab. That works eventually, of course, but it isn’t very efficient. So looking at all the existing bioimaging networks that I have been actively involved with, it made perfect sense to set

Claire Brown

up a user group for expansion microscopy, within that framework.”

“I think about five or six years ago I began to realise how powerful my professional network is. I realised my scientific knowledge is valuable but if I don’t know the answer to a question or how to apply a new technique, I do know someone I can contact who can answer the question and provide a springboard, pointing me to the best resources and collaborators. I also hope people in my network will reach out to me if I have knowledge or information they need. This way we can really learn from one another and put that knowledge into practice, accelerating scientific progress.”

Natalie Woo, a Master’s Student at McGill working under Claire’s supervision, is leading the experimental efforts to establish the expansion microscopy technique in the Brown lab with plans to disseminate it to the broader McGill community through the ABIF. She sees how the impact of the COVID-19 pandemic and widespread acceptance of the virtual meeting platform has paved the way for the success of online user groups such as theirs.

Natalie says: “It definitely helps that we are all really used to this way of doing things, so we can meet very quickly and effectively. I think before, there was maybe a bit of a tendency for people not to make the effort to come to virtual meetings, with a preference for in-person meetings where there were some drinks and snacks on offer!”

The inaugural Expansion Microscopy Group Meeting took place in June 2022, and proved to be a huge success. So much so, in fact, that the followup interest from microscopists working across the globe posed a new problem for the organisers.

Natalie explains: “At first, we were wondering

if people would even be interested in logging on, but we had so many emails from people around the world that we realised we couldn’t come up with a suitable time for everyone to meet regularly. So we decided that we should have a counterpart meeting in a different time zone, and that is where our colleague, Gabriela Segal at University of Melbourne in Australia stepped in. Meetings are run one month on time that is convenient for the Americas and Europe, and the next month for the Asia Pacific. This allows us to stay in parallel with one another as well as moving forward together as one group. The meetings are now well established, but we know there are still a lot of people we haven’t reached, and we want to make this as global and international as possible.”

Claire adds: “We are currently building a database of resources - for example, protocols, recommended reagents, key publications and a discussion board. We are trying to make everything as accessible for people as possible.”

Speakers and attendees at the meetings, and those contributing to online discussions, come from a very wide range of backgrounds and levels of experience. It’s one of the strengths of the growing community, according to Natalie.

She says: “There are a lot of people doing expansion microscopy who all have their own experiences, and that is the beauty of it.We want to bring in speakers of all levels of experience, as long as they want to talk about the research.”

Another recurring theme of the talks given so far, is an openness and willingness on the part of the speakers to shine a light on ‘what didn’t work’ in their experiments. In an emerging field, the opportunity to hear such cautionary tales can help researchers rapidly adopt the technology.

Claire explains: “The whole idea is to help people get started faster, and I have been really impressed that the experts have been so willing to help and share, and get other people going.

Natalie Woo

“If you’re invited to a seminar, it’s natural to want to showcase all your successes, but it might be more useful for the community – and really add value - to talk about things that didn’t work and how you solved these challenges. There is a real need to change the culture around science and encourage researchers to talk about challenges and difficult experiments, and with the national and international bioimaging community, I think there is a real opportunity to do that.”

Natalie adds: “With this technique there are a lot of little things that can go wrong. For instance, there’s a photo-bleaching technique I have been trying out recently, and without some of the insights I have gained from being part of this group, I wouldn’t have made the progress I have made.”

Still less than a year old, the group has already made huge strides in a short space of time, but what’s

next on the horizon?, and how might things look in another 12 months?

Claire says: “I think we are still in the exploratory phase. Right now everything is sharable in the group, but as things grow, the question of how to make information available to the broader research community is something we’re looking into.”

“This whole experience has been such a great way to see how a young student like Natalie can successfully start her master’s project with the expansion microscopy user group acting as a real springboard. Our hope is that many members of this user group will have a similar experience and we will move scientific discovery forward further and faster through this vibrant community.”

Read more about the Expansion Microscopy User Group

Owen Morton

Submit to infocus

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

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.

- an online microscopy lecture series

Stefanie Reichelta, Alex Sossickb, Nick Barryc, Alessandro Espositod , Georgina Fletchere, and Kirti Prakashf,g

a. CRUK Cambridge Research Institute, Robinson Way, Cambridge CB2 0RE, UK

b. Wellcome Trust and Cancer Research UK Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge CB2 1QN, UK

c. MRC Laboratory of Molecular Biology, Cambridge, CB2 0QH, UK

d. Centre for Genome Engineering and Maintenance, College of Health, Medicine and Life Sciences, Brunel University London, Uxbridge, UB8 3PH, United Kingdom

e. BioImagingUK and the Royal Microscopy Society, 37/38 St Clements Oxford OX4 1AJ

f. Integrated Pathology Unit, Centre for Molecular Pathology, The Royal Marsden Trust and Institute of Cancer Research, Sutton SM25NG, UK

g. Correspondence email: kirtiprakash2.71@gmail.com, @kirtiprakash25 (Twitter handle)

Imaging ONEWORLD covers sample preparation, labelling strategies, experimental workflows, and ‘how-to’ image and analysis. The webinars examine inspiring new scientific ideas and facilitate collaborations, with speakers also available for questions and answers. The organisers, core facility and research staff from the MRC-LMB, ICR/Royal Marsden Trust, and the University of Cambridge can also continue the discussion and provide advice on your imaging projects.

In early 2020, everyone was thrown out of their routines, environment, and ways of living. In the run-up to the lockdowns, we scrambled to ensure our students were safely returning home, help our families, colleagues and local communities. Immediately, it also became apparent that imaging labs and facilities would be very challenging to manage during the pandemic. We held meetings to plan how to keep colleagues and students safe while using microscopes, eyes onto eyepieces, hands on focusing wheel in small windowless rooms.Then we installed plastic shields, provided gloves, gel stations, face coverings, and reorganised facilities to adhere to the two-metre distance rule.

Eventually, we also had time to sit down, think, talk

and reflect. With labs momentarily shut, lectures and conferences cancelled, these ruminations gave birth to Imaging ONEWORLD - an online microscopy lecture series that even now – returning to normal life - still provides a valuable educational resource for students and professionals in the broad field of microscopy. We were motivated to connect the British and international communities, to provide high-quality content to anyone, anywhere in the world with an internet connection.

The name of the lecture series is in reference to the ‘One World: Together At Home global broadcast & digital special’ aired on the 17th April 2020 in aid of frontline healthcare workers and the WHO, starting with lady gaga performing ‘Smile’ at her home

studio [Ref: www.globalcitizen.org/de/connect/ togetherathome].

We kicked off on 11th May 2020 and we were amazed to see people joining from so many countries with numbers growing week after week. Suddenly, the community came together in a new virtual space, to speak about ideas, discuss projects and teach each other.

Our lecture series covers a wide range of topics related to microscopy, including the history and development of microscopy, different types of microscopy techniques and applications, methods for sample preparation, data analysis and interpretation. The lectures are presented by expert microscopists of any career stage, who provide insights and realworld examples of how microscopy is used in various fields such as biology, medicine, materials science, and more. Some of the speakers are available after the talk on social media sites like Twitter for questions and answers and to continue the discussion and advice on imaging projects.

Another exciting element of Imaging ONEWORLD is our popular quiz! The quiz helps to revise the lecture content in a convivial and informal way, and to stimulate engagement of people in the follow-up Q&A session. With on-site back to normal, IOW with RMS was concluded earlier this year. However, the invaluable microscopy resource is available via the RMS YouTube channel and IOW Twitter page.

Of course, we are all grateful that we could resume in-person meetings. The pandemic has been a tragedy but has also left a legacy of communitydriven initiatives. Our modest contribution is the RMS Imaging ONEWORLD. We are proud that Imaging ONEWORLD has been a free and an inclusive discussion forum which helped in keeping the imaging community connected. Although we are not just promoting and supporting light microscopy, maybe a line from the poem by Amanda Gorman, read at the inauguration during Joe Biden’s swearingin, still seems quite relevant: For there is always the light, if only we’re brave enough to see it, if only we’re brave enough to be it.

Imaging ONEWORLD key links:

Youtube channel:

https://www.youtube.com/watch?v=tIyls9WhMS 0&list=PLoemaChWEBWFj55504_1PywDUi7Z GIRLP

Twitter community:

https://twitter.com/i/ communities/1584932370700025856

Earlier talks (before 9th November 2020) were hosted on the University of Cambridge video archive and can be viewed here:

https://sms.cam.ac.uk/collection/3229623

The founding organisers (clockwise) - Kirti Prakash, Stefanie Reichelt, Nick Barry, Alex Sossick, and Alessandro Esposito.

Linkam publishes new microelectronics whitepaper to support customers in electrical testing

Linkam has released a new whitepaper to support customers in the fast-growing field of electrical and electronics testing. The whitepaper focuses on different applications of electronics testing, providing a thorough examination of the system set-up and results in a useful resource for electronics testing users who are looking to see different electrical testing methods in action, and learn more about the various types of equipment available

One area the whitepaper covers is research undertaken at Swansea University, where researchers are taking temperature dependent measurements on organic photovoltaic (PV) cells, to better understand how these cells work. The insights are of great benefit to the sustainable energy industry and provide a route for manufacturers to design and develop high-performance solar cells that can help secure and maximise the sustainable energy sources of the future.

The whitepaper also showcases the different applications of ferro- and piezoelectrical testing,

and how temperature control offers researchers the opportunity to study the effects of temperature variation on these materials. Through the use of customised testing apparatus, researchers can now advance their studies in ferro- and piezoelectricity, which may open the door to further applications, such as developments in infrared detection and thermal imaging, and new testing methods in the future.

Dr Andrew Davies, Applications Scientist at Linkam, comments: “Our new whitepaper comes at a time where electronics as an industry is evolving at a rapid pace, and new research and testing methods must keep up with demand so that technology can continue to advance. We hope this whitepaper will be a helpful resource for customers and researchers who undertake electrical testing, and we’re excited to see what the future has in store for electrical applications.”

www.linkam.co.uk

EM Resolutions Ltd has a new home in Keele University Science and Innovation Park

With great excitement, EM Resolutions have now moved into a new production laboratory and office space in the Science & Innovation Park at Keele University in Staffordshire.

Keele University is a student focused university, ranked by students as No. 1 in the UK but has thriving links to industry with over 50 businesses based at the Science & Innovation Park with a specific spotlight on driving growth through innovation.This is the perfect fit for a company with the mindset of innovating to deliver for customers.

This is a fantastic opportunity to put down roots at a site with brilliant facilities, access to the university’s research equipment and talent and be surrounded by a range of fantastic, innovative businesses.

It would be remiss not to acknowledge previous address changes in recent years, mainly due to a shift to home working during Covid however this new space marks the start of the next chapter for EM Resolutions. In addition to the superb opportunities that Keele University provides, there is greater office space and a brand new production laboratory to better serve customers through increased capacity without sacrificing all-important quality.

www.EMResolutions.com

Linkam presents cryo-plunging techniques for automated vitrification of cryo-electron microscopy samples at Microscopy Conference 2023

Linkam Scientific will present a new cryoelectron microscopy (cryo-EM) research paper at the Microscopy Conference 2023 (MC2023), Darmstadt, Germany: ‘New principle and robot for vitrification of cryo-EM samples with controllable sample thickness and real-time optical inspection.’

Led by Linkam’s senior research scientist, Dr. Michael Schwertner, the presentation is taking place at 12:15pm (CET) on 1 March and will be part of the MC2023 Instrumentation and Methods session, within the Development of Cryo-EM Instrumentation and Techniques track.

The paper explores how, despite recent progress in cryo-EM instruments to support automated sample loading and imaging, the routine preparation of cryo samples still presents a challenge. Current cryo-EM methods are held back by the speed and quality of sample preparation, because of the difficulty of creating a uniform film, and a more robust and reproducible method has long been sought by the research community. The paper presents that

utilising suction rather than blotting paper to adjust the film thickness before plunging can improve sample preparation, by removing the potential for the blotting paper to create a challenging environment inside the high humidity chamber. It also gives access for real-time imaging that is used to assess sample conditions and to determine the best moment to trigger plunging.

Dr. Schwertner shares an insight into his presentation: “By comparing traditional cryo sample preparation methods with our new approach of sample application via dipping, followed by film thickness adjustment via suction, researchers are able to optimise cryo-EM sample preparation. Plunging a sample grid into vitreous water delivers structural preservation of biological samples at atomic resolution.We envisage that future decisions on the precise moment to plunge the sample will be automated via an artificial intelligence (AI) vision system.” www.linkam.co.uk

DriveAFM wins Wiley Analytical Science Award 2023

easy to use. It is also a confirmation that after only two years in the market, the DriveAFM is now established as one of the most sought-after AFMs in the very competitive high-end sector.

We are pleased to share that Nanosurf’s DriveAFM was selected by Wiley’s readership as the winner of the 2023 Wiley Analytical Science Award in the category Spectroscopy and Microscopy.

For Nanosurf this is not only a reward for the outstanding work of our development and engineering teams, who designed and built this instrument with the goal to create a stable, highend AFM for diverse types of research that is still

At the award ceremony Dr. Jonathan Adams, who was involved in development from the very beginning, expressed what an honour it is to have been awarded this 1st place: “Developing the DriveAFM was a fun and challenging project, and a real team effort, because of the ambitious goal to achieve the highest level of AFM performance while implementing novel features like photothermal excitation and full motorization in a tip-scanning AFM. We had the freedom to imagine things from the ground up, and we are all really proud of the result and the recognition given to us by the jury and the readers of Wiley.”

www.nanosurf.com

Left to right: Jonathan Adams, Dominik Ziegler, Christian Bippes, Andreas Lieb, Lukas Howald, Lukas Dolfo.

Investigating the properties of meat alternatives and the sensory experience

Currently around 14% of UK adults (7.2 million) are following a meat-free diet, with 8.8 million pledging to go meat-free in 2022. Eating meat alternatives comes with resistance, as mimicking the taste of meat and matching consumers’ multi-sensory experiences has proven challenging. While meeting sensory expectations is one important facet, the nutritional value is another crucial factor.

Mechanical testing is a characterisation technique used in product design and manufacturing. Reading Scientific Services Ltd (RSSL), is a contract research organisation that provides consultancy to the food, drink, and consumer goods sector to help the industry develop and launch new products. Here, the RSSL Food R&D team compared the properties of chicken with new plant-based alternatives developed by a meat-alternative brand using a Linkam Modular Force Stage (MFS), a mechanical testing device, to measure parameters relating to sensory experience.

For the sample preparation, the team used chicken breast and three-plant based chicken products, two soy protein-based and one soy and pea proteinbased. The samples were prepared on a supporting frame and tensile grips were used to hold the sample assembly. The samples were cut to similar sizes from the cooked foods, approximately 20

x 5 x 5 mm. A Linkam MFS was used for tensile and compressive testing at room temperature and pressure. To capture images simultaneously to the experiment, the stage was incorporated with an optical zoom microscope.

From the tensile data, the RSSL scientists were able to create force-extension and force-time graphs from both the tensile and compressive experiments.

Results

All samples failed in a similar way, with fibres breaking under tensile testing. Despite the meat alternatives being close replications of chicken, it was only the soy based protein II that had similar fine fibres with close similarity to chicken.

Conclusion

Understanding the textural properties of plant based alternatives is key to their future success. Creating a comparable multisensory experience will influence how manufacturers create a meat-like plant-based alternative. The MFS provides crucial insights into how meat alternatives compare to meat and can be used to guide future development of superior meat mimics.

www.linkam.co.uk

Have You Found an Image in Your Image?

It’s that time of year again, when CoolLED is accepting entries for its imaging competition with a difference.The prize is a three-channel pE-300white, which is a powerful and reliable LED Illumination System for everyday fluorescence microscopy, featuring:

• Three controllable LEDs covering the most common fluorophores, from DAPI to Cy5

• Software, digital and manual control

• Sustainable, mercury-free fluorescence illumination

How to win a pE-300white LED Illumination System

For the chance to win, all you need is a light microscopy image containing another image, like the 2022 winning entry “Flame” by Claire Dessalles (Postdoctoral researcher, University of Geneva) – pictured above. If you’ve seen a goblin in your gastrula, a horse in your histology or spotted something else entirely, we can’t wait to see your images! The rules are simple: images must be captured using light microscopy, and the closing date is 31st July. Good luck!

www.coolled.com/competition

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.

New to Agar Scientific: Carb-N-Grids - Carbon FIB Lift Out Grids

These grids are manufactured from pure carbon and show very low background for EDS and spectroscopy. The grids have very high adhesion of beam-deposited platinum, allowing for highly stable attachment of sample lamella.

Carbon FIB grids are ideal for spectroscopic examination and where metals such as copper must be avoided. The unique carbon material is mechanically robust and provides smooth edges at attachment points.

The major advantage of the grids is the Zero-ZeroZero method:

• Zero pre/post alignment due to bent grids

• Zero fluorescing

• Zero waste

www.agarscientific.com/carb-n-grids

Prior Scientific introduces the H189 Motorized XYZ Deck

Prior Scientific, a manufacturer of microscopy solutions and precision optical and electromechanical equipment, today announced the launch of the H189 Motorized XYZ Deck. Prior Scientific’s new motorized XYZ Deck is the latest in the Deck series from Prior. As a natural progression from the original ZDeck the new XYZ Deck is purposely designed to match the increased size and versatility of the latest generation of 2P/3P/ Multiphoton/Confocal microscopy systems giving researchers the flexibility to utilise all the potential of the added sample space and microscope power of these new microscopes.

The XYZ Deck incorporates two removable shelves with an impressive 25kg load lifting capacity. The removable shelf feature of the XYZ Deck greatly facilitates sample preparation when working with larger specimens or preparing complex experiments. Oversized breadboard deck plates make the XYZ Deck the optimal solution for a large range of sample types from whole animal

invivo to slides and petri dishes. Compatible with Queensgate Piezo models, the XYZ Deck is compatible with most imaging software and ideal for use with manipulators, probes or other sensory devices. With 50mm of motorized in XY and Z travel and a sample height adjustable from 79.5mm to 410mm, the XYZ Deck is a versatile and robust system that will prove to be a valuable addition to your system configuration.

www.prior.com

New piezo objective scanner offers market leading accuracy and resolution

Nanopositioning expert, Queensgate, has announced the launch of the Nano Scan OP800 piezo objective scanner. Its unique mechanical design together with integral capacitive feedback sensors offer users market-leading positioning accuracy, while its stiff construction allows the standard selectable tuning pre-sets to support a range of objectives from 150 g to 500 g. This makes it highly versatile for use in a wide range of applications in life sciences and industry.

The OP800 offers 800 µm of closed-loop travel range for easier operation within the piezo focus range without adjusting the sample. It also allows deeper Z-stacks to be completed: ideal for multiphoton imaging techniques. Auto-calibration on power-up automatically offsets the range to correct for load and environmental changes to maintain the full 800 µm travel.

It is flexure-guided with built-in capacitive position feedback. This provides high linearity of motion and

market-leading repeatability of just 5 nm, ensuring consistency between multiple Z stacks. Small rotational errors help ensure the full field of view stays in focus.

Images are captured rapidly thanks to the fast stepsettle times. Constant velocity mode operation allows rapid precision ramping to form very quick image stacks over long ranges. This is several times faster than conventional position control can achieve.

As is standard with Queensgate stages, the OP800 offers true plug-and-play capability without the need to match controllers. Both digital and analog interfacing are easy to set up, while the lownoise, high-powered digital electronics allow rapid stepping. The OP800 is suitable for use in life sciences, materials and semiconductor applications, such as optical sectioning, surface analysis, wafer inspection and scanning interferometry.

www.prior.com

Meet Dragonfly 600, Oxford Instruments’ New Flagship Confocal Microscope

Oxford Instruments Andor, a world leader in scientific imaging solutions, has announced the launch of Dragonfly 600, further strengthening its award-winning confocal microscopy portfolio. Dragonfly 600 introduces three groundbreaking features; a new total internal reflection fluorescence (TIRF) modality: Borealis TIRF (B-TIRF), a high-power laser engine, and a unique design of 3D Super-Resolution module which retains parfocality across all imaging modes. These technical innovations, combined with innovative software, extend Dragonfly’s capabilities to excel at single molecule localisation microscopy, delivering results with nanometre precision.

This new flagship product also maintains the benefits that position Dragonfly as the leading confocal microscope in the market: it is 10 times faster than point scanner confocals, it images very deep into thick organisms, and it allows users to

and contribute to high throughput for emerging applications such as mapping brain transcripts or phenotyping cancer gene expression. Cell biologists will be able to uncover the ultrastructural organisation of organelles or cellular membranes with resolutions an order of magnitude greater than the diffraction limit of light.

Flexibility and ease of use are enabled through open-source Picasso software, adapted for seamless data exchange, and enabling localisations with nanometre precision. Meaningful statistics and beautifully rendered images are then delivered through Oxford Instruments’ market-leading image analysis software, Imaris 10.0.

Dr. Bernhard Goetze, Oxford Instruments Andor's Strategic Head of Life Sciences for Product Management, commented: “The Dragonfly 600 will allow the researchers to access superresolution microscopy with confocal imaging. It enables cell biologists, developmental biologists, neuroscientists, and other life science researchers to acquire super-resolved images inside thicker tissues, which were not

Brand New AFM Image Taken Inside an OptiCool Optical Cryostat

A new Atomic Force Microscope image was recently taken inside a Quantum Design OptiCool optical cryostat, at 200 K and 6.6 T. This image demonstrates the low noise and low acceleration environment inside the OptiCool. The AFM is part of the scanning optical nearfield microscopy (s-SNOM) built by Mengkun Liu and his research group at the Department of Physics and Astronomy at Stony Brook University, New York.

“We were looking for a closedcycle magneto-optical cryostat that is capable of performing AFM and s-SNOM in high magnetic field”, said Mengkun Liu, Stony Brook University. “OptiCool proved to be the system with the best stability which provides sub-nm vibrations for cyrogenic scanning probe measurements while still offering excessive access to external light. This brings us infinitely new possibilities for our research. Our lab now hosts two OptiCool systems - one for infrared magneto-SNOM and one for terahertz magneto-SNOM.”

Mengkun Liu is an associate professor at the Physics Department of Stony Brook University. His research interests include physics of correlated electron systems, two-dimensional materials, infrared nano-optics and ultrafast time domain THz spectroscopy. Prizes include NSF Career Award (2021) and Seaborg Institute Research Fellowships at Los Alamos National Lab (2009).

The OptiCool® by Quantum Design is an optical

cryostat using an innovative design that puts the sample volume in the heart of your optical environment. A custom 3.8 inch bore, split-coil, conical magnet offers fields perpendicular to the optical table up to ±7 tesla. The highly integrated design means, even with a magnet, your sample isn’t buried inside a large cryostat, far away from the optics. Seven side optical ports and one top optical port allow for optical access to your sample from a wide array of directions.

Dr. Luke Nicholls, Technical Sales Manager at Quantum Design UK and Ireland, commented: “At Quantum Design we are proud that the innovative OptiCool Magneto Optical Cryostat can enable our customers to push boundaries and produce ground-breaking research. Congratulations to all our customers who make the OptiCool user community so successful.”

www.qd-uki.co.uk

Imaris Launches 10.0, an AI-powered Software For Fast and Accurate Detection of Neurons, Microglia and Blood Vessels

Oxford Instruments - Andor has released Imaris 10.0, the latest version of its market-leading microscopy image analysis software. Imaris 10.0 revolutionises the well-known and trusted Filament Tracer, which has been used by the neuroscience community for over 15 years, leading to almost 700 scientific papers.

The new 10.0 Filament Tracer combines the versatility of an Autopath intensity-based method with a unique machine learning approach, providing the most flexible tool on the market for tracing a multitude of neuron types.

are separated from the background.

What differentiates Imaris 10.0 from other packages is the enablement of automated distance measurements between filamentous structures and other stained objects. It’s been measured to be faster by a factor of 5 than its predecessor and easily handles much bigger datasets.

Meredith Price, Software Manager for Oxford Instruments – Andor, says: “The advantage of Imaris 10.0 Filament tracer over the other popular approaches lies within its multi-purpose robust methods, which enable creation of proper filamentous models for various types of biology datasets, from neurons to tiny blood vessels. The fact that no prior training is needed means the tool can be used in a multitude of labs.

This Filament Tracer can resolve various structures, such as neurons, dendritic spines, microglia, blood vessels, endoplasmic reticulum*, and microtubules* within minutes. Creating filament models is very easy, even for non-experienced users, because they are guided via precisely designed wizards with the result previewed after each step. By following the wizard instructions accompanied with a few paintbrushes along the structures of interest, they

“The latest release greatly improves the speed of calculation and visualisation of the detected models, which is one of the most important factors for scientists regarding the size and complexity of datasets they are willing to analyse.”

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. https://andor.oxinst.com/

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.

Meet the Staff: Nick Cameron, Sponsorship & Events Assistant

Nick plays a key role in the RMS events team, bringing in the allimportant event sponsorship and helping to keep the society’s programme of meetings, conferences and courses ticking over. From initial contact with venues and site visits, to securing the sponsorship bookings, and finally attending the events and meeting the sponsors themselves, Nick is involved at all stages of the process. Since joining the RMS in March 2022, he has also been filling in as the main point of contact for Corporate Members, while colleague Chloe Goode is on maternity leave.

first fully-in-person conference since the onset of the Covid pandemic – Frontiers in BioImaging 2022 –was his very first event for the Society.

He recalls: “It was great to see all the bookings start coming in, and before long we were sold out. It was really nice to be part of our first proper meeting since the pandemic and to meet all the sponsors – all the people I had been emailing in the months beforehand.

“We’re obviously now working hard on preparations for mmc2023. That takes a lot of planning but we’ve also got quite a lot of events going on at the same time. We had three events in January and then there are three more in April.We’ve also been to Liverpool to have a look at the venue for elmi2024, and to Warwick for an event happening in September.”

With so much going on, how does Nick stay on top of each project, and what makes for a successfully delivered event?

Nick’s arrival at the RMS coincided with the return to in-person meetings in mid-2022. In fact, the Society’s

He says: “It’s really important to stay on top of everything that’s going on, and making sure everyone has got the right information. That communication is so important, and it is always good to go and visit the venue in advance and build up a good relationship with the venue. Once you’ve got that in place, you feel

Family time: Nick on holiday with wife Lisa and son, Jack.

more confident that things are going to run smoothly.

“I’m very excited about mmc in the summer, as I have worked on a few small exhibitions but never anything on this scale. We’re working closely with all the exhibitors, and we’ve invited them all to come and view the exhibition space. It’s been a while since we last hosted mmc in 2019, so we want to make sure people have the opportunity to come and have a look, and to ask any questions they might have. Hopefully we can deliver a really great event and get plenty of people visiting the stands.”

Nick has many years’ experience working in the hospitality and catering industry, having worked in management roles at hotels across England, and most recently, in events management at the Kassam Stadium – home of Oxford United FC (Nick’s number one team – of which more later).

At the age of 17, he began his career as a general assistant in a small hotel in Oxfordshire. From there, he moved to a bigger hotel in Lyme Regis on the English south coast, now in an assistant managerial position. There followed a series of roles in conferencing and events for a hotel chain with venues in Oxford and Leeds.

Nick says: “At Leeds we were doing dinners for up to about 500 people. I remember on my first day, we had the Leeds Rhinos rugby league team’s end of season

dinner, which was quite an event!”

From 2010 onwards, Nick worked at the Kassam Stadium, initially as Assistant Operations Manager looking after conferences and events, including corporate hospitality for Oxford United’s home matches.

“There were about 30 executive boxes, and we had around 20 agency staff looking after them on match days. Then I moved into more of an office role, booking the non-match day events, mainly because my son had just been born and the working hours fitted better around family life.”

With the Covid pandemic hitting the events industry particularly hard, Nick’s office closed down and he was furloughed for over a year. By the time he returned to work, the conferencing area of the Kassam had been turned over to an enormous vaccination centre. It was ultimately the signal for Nick to jump ship – and what brought him to the RMS.

He says: “By the time we started taking bookings again, we had very limited space available. Covid vaccinations were still taking place every day in our two largest conference rooms, and we could only offer small meetings. A lot of my colleagues had left during Covid, and there was no certainty over anything.”

Having now been at the RMS for almost a year, what

Spoils of victory: Nick celebrating a league-title win for his team, Steeple Aston.

Nick Cameron

have Nick’s impressions been about the Society, and his new working environment?

“From what I have noticed in my time here, it seems that the microscopical community is quite a tightknit community”, he says. “I think the RMS staff really understand the importance of that.They care about it and that makes a real difference.

“I didn’t know anything about microscopes, so I was a bit worried about that to start with, but I’ve been getting to know more about the different types of sciences, and which companies are linked with the different types of microscopy. You start to get a feel for what people are going to be interested in, which is really useful.”

“Everybody has been really helpful, and the transition has been a lot easier than I was expecting – even with more working from home, it has been easy get to know people.”

Nick lives in Bicester with his wife Lisa, and eightyear-old son Jack. He also has an older stepdaughter, Georgia, who moved out of the family home a couple of years ago.

Among Nick’s interests, he is a keen cook, never happier than when rustling up a few culinary treats. He says: “Working in hotels for years, I managed to pick up a few tips in the kitchen, and my wife also used to be a chef. We share the cooking, but I like doing a nice roast dinner on a Sunday.”

However, Nick’s true and lifelong passion is football – a sport he has played since childhood, representing various local teams in Oxfordshire – including nonleague outfit Banbury United at one stage. He is a Committee Member at local league side Steeple Aston FC, and still occasionally plays for them on Saturdays – either at full-back or centre midfield. He also regularly plays for nearby Chesterton Vets on Sundays. As if that wasn’t enough, he somehow finds time to coach his son Jack’s team, Bure Park Under-8s.

Saturday afternoons (at least, every other Saturday) can also only mean one thing for Nick – a trip with Jack to watch Oxford United at the Kassam.Through thick and thin (mostly ‘thin’, if we’re being honest) Nick will be up there in the stands, cheering on the mighty Yellows.

In 2011, he even got down on the pitch to play alongside some of his heroes, after winning a ballot to take part in a testimonial match for former United legends David Langan and Joey Beauchamp. Nick ended up on the losing side that day, but not before getting on the scoresheet in front of around 2,000 spectators. As proud moments go, it’s not a bad oneand we even have the photo to prove it!

Guiding hand: Nick on coaching duty with Jack, after a recent tournament win.

Pick that one out!: Nick turns to celebrate after scoring at Oxford United’s Kassam Stadium.

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

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

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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.

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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.

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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).

Figure 1. Width of figure or table confined to one column.

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

Two column/full page width, 135mm

ATLAS 5 micron MicroXRF

• The only benchtop system with - a perfectly circular 5 micron spot size

- a total of 4 independent detectors and the largest total detector area of 600mm2

- the largest chamber size in the Atlas X version

- a highly intuitive, quantitative software in the Iridium Ultra

• The only system that can be operated with the doors open, X-ray o

• The system can also measure Carbon and other light elements

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