Tecan Journal Life Sciences and Partnering Business
Sunrise™ in Mongolia pages 22-23
Flexible automation for next generation sequencing library preparation pages 12-13
Researchers at Nagasaki University develop method for antemortem CJD detection pages 16-17
Freedom EVO® in Evogene’s Biolab runs 13 cloning related procedures pages 24-25
CEO WELCOME TECAN JOURNAL 3/2011
Your trusted partner in a turbulent market. As international markets continue to deal with global economic instability, we are beginning to see a recovery in the laboratory automation sector. Recent events have shown that it is important to have reliable partners, whether you are a small academic laboratory or a major pharmaceutical company. With over 30 years of knowledge and experience in the laboratory automation sector, Tecan’s innovative, customer-focused approach means that we are able to deliver the best solution for your needs, however complex they may be. This track record of reliability provides important reassurance for our life science customers, and allows our OEM partners to invest in Tecan technology with confidence, as demonstrated by another recently announced major OEM agreement with a global leader in the diagnostic sector.
In this issue of the Tecan Journal you will find details of some exciting product launches, including the HP D300 Digital Dispenser – the first product from our collaboration with HP to accelerate the drug discovery process – and an updated Infinite® M1000 multimode reader. We also have examples of the flexibility of our instruments, demonstrating how the Freedom EVO® platform’s MCA 384 option is helping customers perform various applications in drug discovery, siRNA screening and the development of novel molecular tools. At the other end of the spectrum, we have an article about Tecan’s involvement in a national health project in Mongolia, providing an interesting look into the difficulties of performing good science with limited resources. We hope you enjoy the issue,
Thomas Bachmann, CEO
Stop Press! Tecan and HP have won the prize for the best new technology at the annual ELRIG meeting with the HP Direct Digital Titration technology (see page 6), based on voting by the 1200 attendees.
Kevin Peters (HP Global Sales Development Manager) and Jessica Merlino (Tecan Product Manager, HP D300 Digital Dispenser) were presented with the Technology Prize by ELRIG Director Stuart Baddeley
CONTENTS TECAN JOURNAL 3/2011
Tecan brings HP Direct Digital Titration to drug discovery biologists
4 Enhanced pipetting flexibility with new MCA 384 Disposable Tips 5 The future of microplate detection has arrived 6
ecan brings HP Direct Digital T Titration to drug discovery biologists
7 Gambarou Nippon! Let’s do our best together Japan! 7 Tecan-sponsored graduate team triumphs in Californian sun 8-9 MCA 384 adapter plate concept and pin tool provide the ultimate flexibility 10-11
The future is smaller
12-13 Flexible automation for next generation sequencing library preparation pages 28-29
14 Injecting new life into environmental science
Pressure Monitored Pipetting supports Swiss BioQuant’s dedication to excellence
15 Automating advances in medical technology 16-17 Researchers at Nagasaki University develop method for antemortem CJD detection 18-19 Relying on automation – new applications for circular dichroism 20-21 Pressure Monitored Pipetting supports Swiss BioQuant’s dedication to excellence 22-23 Sunrise™ in Mongolia 24-25 Freedom EVO in Evogene’s Biolab runs 13 cloning related procedures 26-27 Automation of DNA analysis boosts the fight against crime 28-29 Second generation biofuels – making energy from waste 30-31 Quality matters: audits as a learning platform 31
Leading the Debate
PRODUCT NEWS TECAN JOURNAL 3/2011
Enhanced pipetting flexibility with new MCA 384 Disposable Tips Tecan has launched a new generation of high quality MCA 384 Disposable Tips designed to optimize the performance and reliability of the versatile MultiChannel Arm™ (MCA) 384 pipetting option on Freedom EVO® workstations. The all new range of MCA 384 Disposable Tips has been extended to include a 15 μl tip size alongside the existing 50 μl and 125 μl sizes, providing high quality, reliable pipetting across a broad volume range.
New 15 ml MCA 384 Disposable Tips with filter
All MCA 384 Disposable Tips are now available in our enhanced Tecan Pure and Tecan Sterile purity levels, with filtered versions of Tecan Sterile tips also available in all sizes. Tips are precision manufactured to deliver reliable 96- or 384-channel pipetting, as well as single row or column pipetting, and have a patented design which ensures high accuracy and low CVs even at low volumes, helping to save valuable reagents and reduce maintenance requirements. The new range offers maximum format flexibility, including the use of 1,536-well plates with 15 μl tips, full depth access to 96 deep-well plates with 125 μl tips and use of typical 0.7 ml and 1.4 ml 96 storage tube formats, providing a comprehensive solution to help you meet your automation needs. In addition, each product is supplied with a Certificate of Conformity (CoC) to ensure full traceability and guarantee that the product meets the stated specifications, helping you to meet regulatory and quality documentation requirements.
To find out more on Tecan’s MCA 384 Disposable Tips, visit www.tecan.com/consumables
15 µl, 50 µl and 125 µl MCA 384 Disposable Tips
PRODUCT NEWS TECAN JOURNAL 3/2011
The future of microplate detection has arrived Tecan has recently updated its Infinite® M1000 high-end multimode microplate reader with new features designed to offer the same top class performance for an even wider range of applications. First launched in 2008, the Infinite M1000 uses Tecan’s premium Quad4 Monochromators™ technology to offer exceptional speed, sensitivity and flexibility for academic, biotechnology and pharmaceutical research applications. Already highly acclaimed for its scanning capabilities, the Infinite M1000 now offers luminescence scanning to complement its existing 3D and ultra-fast absorbance and fluorescence scanning functions. Luciferase reporter mechanisms have become commonplace across a broad range
of disciplines, making development of recombinant luciferases and novel substrates an important area of research. This new feature allows the emission spectra of stable luminescent signals to be recorded at 1 nm resolution using the instrument’s emission monochromators, making the Infinite M1000 an ideal platform for luminescence-based assay development. The format flexibility of the Infinite M1000 has also been expanded, allowing Tecan’s patent pending NanoQuant Plate™ for very low volume absorbance measurements to be used alongside virtually all 6- to 1,536-well plate formats. A new double orbital shaking option – moving the plate transporter in a figure-of-eight motion – offers enhanced mixing of reagents and biomolecules, as well as improved suspension of non-adherent cell cultures. For adherent cell cultures, the introduction of an Optimal Read (OR) function has further enhanced measurement reliability in bottom reading fluorescence intensity mode, by performing multiple, spatially separated measurements to achieve maximum illumination and sensitivity. The new Infinite M1000 also features a redesigned injector module, offering up
to two injectors for precisely controlled pipetting for triggering fast kinetic reactions and bulk dispensing. This state-of-the-art option is available with various syringe sizes and features a tilting reagent carrier to ensure low dead volumes and minimize wastage of substrates and buffers, as well as a bulk dispense function to eliminate tedious pipetting steps for 6- to 384-well plates, saving you both time and reagents for plate-based kinetic studies. The modular, upgradeable nature of Infinite series readers means that customers who purchase the new Infinite M1000 will also be able to benefit from the soon to be launched AlphaScreen® option, making the Infinite M1000 a truly flexible solution to meet your changing laboratory needs. To find out more about Tecan’s Infinite M1000, visit www.tecan.com/infiniteM1000 NOTE: New features are only available for the updated Infinite M1000 (REF 30061442) with V2.0 or higher firmware and V1.8 or higher i-control™ software. For further information please contact your local sales representative. AlphaScreen is a registered trademark of Perkin Elmer, USA.
LIQUID HANDLING & ROBOTICS TECAN JOURNAL 3/2011
Tecan brings HP Direct Digital Titration to drug discovery biologists
The new HP D300 Digital Dispenser is a simple, standalone instrument using HP’s Direct Digital Dispensing technology to rapidly deliver picoliter to microliter volumes of drug compounds.
Direct digital dispensing system allows any dose in any well
Available exclusively through Tecan’s Life Science Business, this innovative device is the result of an agreement between Tecan and HP to commercialize new automation solutions for drug discovery, and can cut titration times to just minutes. The HP D300 provides fast and reliable performance across a large dynamic range, eliminating the need for slow, error-prone and wasteful serial dilutions. Using disposable dispenseheads to help avoid the possibility of cross-contamination, the instrument offers non-contact dispensing from 13 pl to 5 μl, delivering any dose to any well and virtually eliminating waste of valuable compounds. It combines walk-up convenience, ready-to-run performance and flexible, intuitive experiment design software to increase research productivity, reliability of results and the speed of drug discovery workflows.
Wendy Lauber, Head Product Management for Liquid Handling and Consumables at Tecan, commented: “The pharmaceutical industry is recognizing a need for change, and there is a growing trend towards benchtop automation solutions. New technologies such as Direct Digital Titration can offer fresh solutions to entrenched challenges, helping to improve productivity and accelerate the discovery of better drugs.” The HP D300 Digital Dispenser is available to order now in Europe and North America. To find out more about Direct Digital Titration, go to www.tecan.com/digitaltitration
The HP D300 Digital Dispenser is available to order now in Europe and North America. To see the HP D300 Digital Dispenser in action, visit us at: 20 - 22 Sept 2011
11 - 13 Oct 2011
23 - 27 Oct 2011
02 - 04 Nov 2011
Discovery on Target
CORPORATE NEWS TECAN JOURNAL 3/2011
Gambarou Nippon! Let’s do our best together Japan! In the aftermath of the deadly earthquake, tsunami and radiation threat in March, there was widespread fear that this disaster would also impact on Japan’s national economy, with the country being left behind in the global market. For the first couple of months after the disaster, most of the tradeshows, festivals and concerts were cancelled, with International BioEXPO – the annual life sciences trade show held in Tokyo in June – among those threatened. Making a success of this show had special significance for Tecan Japan, helping to create a positive impression and further encouragement for the Country’s industry, and we are pleased to report that the show was highly successful, attracting similar visitor numbers to last year. “The devastation caused to the earthquake and tsunami-hit area is huge, and far beyond any description,” commented Miyako Sakai, Marketing Communications at Tecan Japan. “With more than 20,000 dead and missing, many lost their loved ones, houses and jobs. This tragedy has reminded the Japanese
people of their fortitude in the face of adversity and of the strong ties we have with the rest of the world; we appreciate very much the support and prayers we have received from across the globe. Life goes on and we are hanging in there with our favorite slogan: ‘Gambarou Nippon!’ Let’s do our best together Japan!”
Tecan and its employees worldwide have contributed a total of CHF 50,000 to the relief effort through Swiss Solidarity, a foundation that raises funds for humanitarian projects and works closely with the SRG SSR (Swiss Broadcasting Corporation) and relief organizations around the world.
Tecan exhibits at International BioEXPO 2011
Tecan-sponsored graduate team triumphs in Californian sun management skills within the life science sector. The TMP is the capstone of the MBS program, and is designed to encourage innovative, application-focused thinking as multidisciplinary teams of postgraduate students work with industry figures to solve real problems using novel and existing technologies. TMP project team
Tecan is pleased to announce that a Company-sponsored team has won the Keck Graduate Institute (KGI) Team Masters Project (TMP) Collaborative Excellence Award, part of the Institute’s Master of Bioscience (MBS) degree program focusing on business and
Each year, leading companies from throughout the life sciences industry are invited to sponsor individual teams, helping to investigate how cutting-edge technologies could be applied to novel or previously unexplored market sectors. This year’s Collaborative Excellence Award was won by students Jessica Chang, Neelima Kumar,
Alice Lai, Jessica Lin, Sergio Sanchez and Jeffrey Weiss, who worked with Marc Feiglin and Tony Mamone from Tecan to explore new areas for the application of Tecan’s technology portfolio. Craig Adams, KGI Faculty Supervisor for the team, commented: “This was an exceptional team; they worked closely together while maintaining their analytical objectivity and adding their individual perspective into each market analysis. Finally, Marc and Tony facilitated such a great project that we are thrilled Tecan has chosen to sponsor another TMP during the 2011-2012 academic year.”
LIQUID HANDLING & ROBOTICS TECAN JOURNAL 3/2011
MCA 384 adapter plate concept and pin tool provide the ultimate flexibility Flexibility is vital to researchers at the University of Konstanz, who rely on Tecan’s MultiChannel Arm™ (MCA) 384 and Freedom EVO® liquid handling platform for proteinand cell-based assays.
Scientists in the Biology Department at the University of Konstanz, Germany, are developing novel molecular tools to help aid the investigation of cellular processes. Professor Thomas U Mayer explained: “We are investigating how small molecules affect protein activity, and how they may be used to activate or deactivate this activity in living cells. Our main interest is chromosome segregation during mitosis and, as small
Stefanie Selent, technical assistant in the screening unit, with the Freedom EVO
molecules which exhibit the activity we need are not available, we have to create them ourselves. We perform a variety of proteinand cell-based assays, using fluorescencebased techniques to follow the process of interest. Once hits are identified, we verify their specificity and identify the target, then use them as probes to study chromosome segregation in mammalian cells.”
LIQUID HANDLING & ROBOTICS TECAN JOURNAL 3/2011
The Freedom EVO in the Biology Department at the University of Konstanz
The department depends on a Freedom EVO platform and the versatile MCA 384 option to perform its screening assays, allowing reliable and reproducible pipetting of very low volumes. Thomas continued: “We mainly work in 384-well format – but we also use 96-well plates for RNAi-based screens – and so wanted a highly flexible system in order to switch assays when required. The throughput offered by the MCA is very impressive, but our focus is more on flexibility because we work on a small scale. When first looking at automating this process, we were impressed by Tecan’s approach. Tecan engineers came to our laboratory and gained a thorough understanding of our workflow requirements, then designed a workstation that was tailored to our specific needs, taking
advantage of the MCA 384 option for a variety of functions.” “This high level of flexibility is very important in academia, and the Freedom EVO offers us reliable walkaway operation,” Thomas concluded. “The MCA 384’s adapter plate concept allows us to automatically switch between 384- and 96-well pipetting, and its flexibility has enabled us to use our existing 384-well pin tool giving us the capacity to transfer just 50 nl of material. This enables us to use a single workstation to perform a wide range of tasks, and we have been very happy with the system’s performance.” To find out more about Tecan’s MCA 384 option, visit www.tecan.com/mca384
DRUG DISCOVERY TECAN JOURNAL 3/2011
The future is smaller
Merz Pharma has taken advantage of the greater flexibility and large pipetting volumes offered by the Freedom EVO® workstation’s MultiChannel Arm™ (MCA) 384 option to accelerate its drug discovery pipeline, eliminating the need for single concentration analysis in its screening processes.
Merz Pharma is an independent pharmaceutical company based in Frankfurt, Germany, specializing in drug discovery and development for neurological and psychiatric disorders including Alzheimer’s disease, Parkinson’s disease and tinnitus. The Company’s drug discovery pipeline uses a radioligand-based screening strategy to identify candidate compounds which interact with disease-relevant membrane receptors, as Dr Meik Sladek, Section Head, in vitro screening at Merz Pharma, explained: “Our in vitro screening department is responsible for functional and binding studies for potential drug candidates. We use preparations of membrane receptors – either from tissues or transfected cell lines – to identify novel target binding activities, and our entire assay processing workflow is automated using a Freedom EVO workstation.”
The MCA 384 in operation on Merz Pharma’s Freedom EVO platform
“The availability of membrane material has historically been the limiting factor in our throughput, however we have now increased the number of assays we are able to perform per membrane preparation by switching from a 96-well to a 384-well microplate format. To accommodate this change in our workflow we have upgraded our Freedom EVO workstation, taking advantage of the flexibility of the MCA 384 pipetting arm. Our instrument is equipped with a MCA 384 channel head and, by using different adapter plates, the MCA 384 gives us the capability to automatically switch between 96- and 384-well microplate formats. This has given us greater flexibility than before and enabled us to increase the number of assays we are able to perform per membrane preparation.
DRUG DISCOVERY TECAN JOURNAL 3/2011
Tanja Bauer (left) and Sabine Denk preparing Merz Pharma’s Freedom EVO platform for screening
The instrument prepares seven dilutions of up to nine different drug candidates in a 96-well plate, and then converts these dilution series to a 384-well format, allowing us to quickly and easily run each assay in quadruplicate.” “The pipetting volume range offered by the MCA 384 is also very important for our screening workflow. Because our assay involves both filtration and washing steps, we needed a system that was capable of parallel pipetting at least 100 μl per channel in 384-well format. At the time, most automated liquid handling systems using this format were only capable of pipetting
30 or 50 μl per well, but the MCA 384 is able to reliably aspirate and dispense 125 μl per channel, allowing us to perform our filtration assay rapidly and effectively. This change to our workflow has improved productivity and reduced costs, and could not have been achieved without the pipetting flexibility offered by the MCA 384,” Meik concluded. To find out more about Tecan’s MCA 384 option for Freedom EVO workstations, visit www.tecan.com/mca384 To learn more about Merz Pharma, go to www.merz.com
GENOMICS TECAN JOURNAL 3/2011
Flexible automation for next generation sequencing library preparation The Centre for Genomic Research at the University of Liverpool in the UK is routinely using two Freedom EVO® platforms to minimize errors and alleviate the tedium of manual techniques for the production of fragment libraries for next generation sequencing. The Centre for Genomics Research (CGR) at the University of Liverpool, UK, is funded by the MRC and the NERC, and has evolved from a small academic laboratory in 2009 into a facility offering next generation sequencing (NGS) capabilities to academia and industry throughout Europe. The CGR’s core remit is to aid the research community in accessing the full potential of the latest advances in DNA sequencing and bioinformatics methods, and has included collaboration with the University
University of Liverpool, UK
SOLiD sequencing workflow and Tecan solutions DNA extraction
Freedom EVO magnetic bead or vacuum separation
Freedom EVO & Covaris
Freedom EVO 75 & 150
Templated bead prep.
SOLiD EZ Bead™ System emPCR & enrichment
SOLiD 3, 4 & 5500 series
AB has demonstrated scripts for SOLiD fragment library preparation for SOLiD 3 (EVO 75), 4 (EVO 75 & 150) and 5500 (EVO 75 & 150) systems
How Tecan fits into the Roche 454 workflow
Emul. prep. Breaking & amplif.
Sequencing & analysis
GS FLX sequ.
Tecan integrated REM e System • Bead enrichment • Collecting beads • Primer annealing
of Bristol and the John Innes Centre in Norfolk, UK, on a recent high profile wheat sequencing project. In parallel, the CGR has developed an active, internally driven research program that runs alongside its collaborative service activities. The Centre is equipped with a number of different NGS instruments from all major players, including an Illumina, two 454™ sequencers (Roche) and four SOLiD™ systems (Life Technologies). Dr Daryl Williams, Senior Research Associate, explained: “We use various NGS systems because there is no constant requirement – every set of samples requires a different approach and new NGS capabilities are being developed almost weekly! In 2010, we decided to look for instruments to automate the more labor-intensive steps of NGS sample preparation. The manual production of DNA fragment libraries has always been time consuming, with tedious column-based or AMPureTM bead cleaning steps and high potential for error. We looked at several options for automating our workload and it became clear that Tecan systems fitted our needs better and offered more than other systems we looked at. A major selling point for us was the availability of ready-made scripts for the SOLiD sequencers through Tecan’s collaboration with Applied Biosystems (Life Technologies), plus the proven integration of the Roche REM™ e System for 454 emPCR™ enrichment. In addition, the Freedom EVO 150 offers excellent flexibility to add additional NGS sample preparation protocols as new protocols are being released. Other important factors were the large deck size and the option to choose either 8- or 96-channel pipetting. We decided on the 8-channel option to give us the flexibility to upgrade the platform if required in the future, by adding components like multichannel heads or grippers.”
GENOMICS TECAN JOURNAL 3/2011
Christian Bourne, Sequencing Production Research Associate, using the REM e module on the Freedom EVO 75
The Centre’s Freedom EVO 150 platform is specifically set up for production of SOLiD DNA fragment libraries, where automation really helps with clean-up stages prior to sequencing. Daryl elaborated: “Production of libraries using the automated platform means that the entire process is secure, in terms of adding the correct reagents and substrates at the correct times, and this eliminates the sequencing variability that often arose from manual procedures. We can adapt the scripts according to how much or how little of the process we want to automate for a particular batch of samples. Our process starts with quality control (QC) and DNA quantification, followed by shearing – using Covaris, with the potential to automate on our Freedom EVO in the future if workload demands – QC and cleaning of samples either manually or again, if workload demands, on the Freedom EVO platform.” “The workflow includes end repair with DNA polymerase, addition of AMPure beads, AMPure cleaning, manual addition and ligation of barcode adaptors, AMPure cleaning, nick translation and PCR amplification, QC, final AMPure cleaning and size selection to obtain the correct sized DNA (200-300 bp) for SOLiD sequencing. The whole process of SOLiD library construction, which would have taken five days
manually with a medium size batch of 20-40 samples, is now reduced to just two.” Although saving time is important, Daryl stressed that this was not the only advantage of automation for the team: “The most important aspects of automation for us are security and relief from tedious repetitive tasks. This is also true of our Freedom EVO 75, which is fitted with a REM e module and can be run twice a day, to meet the workflow of our 454 Titanium Sequencing System. We break the emulsion, removing oil from the DNA-bound PCR beads, and then the REM e module performs bead enrichment. The workflow is completely automated after the initial set-up, and gives consistency and reliability, saving two to three hours of tedious manual labor twice daily.” Daryl concluded: “Automation for NGS is continuously evolving at the CGR; our first aim was to automate production of DNA fragment libraries for SOLiD sequencing and we have achieved this with excellent flexibility to handle our variable workflows. We have received absolutely amazing support from Tecan, especially through Alettia Elwin, Field
Automation Specialist at Tecan, who modified the scripts to our specific needs, and spent several days checking both platforms to ensure everything was running smoothly. We are now so familiar with our systems that we routinely modify scripts and are well-prepared to cope with larger sample numbers to feed the everincreasing capacity of our NGS systems. We have just taken delivery of the newest SOLiD 5500 system and will soon implement the AB/ Tecan co-developed and tested protocols for SOLiD 5500 fragment library preparation. Next we are looking at automating exome capture protocols from Agilent and Nimblegen. The flexibility the Tecan systems offer is such an important capability for us to keep up with the speed of development of NGS systems.” To find out more about Tecan’s NGS sample preparation solutions, visit www.tecan.com/ngs For more information on the Centre for Genomic Research, visit www.liv.acuk/cgr/ AMPure is a registered trademark of Beckman Coulter, Inc. 454, emPCR and REM are trademarks of Roche. EZ Bead and SOLiD are trademarks of Life Technologies.
components TECAN JOURNAL 3/2011
Injecting new life into environmental science
Picarro is using Cavro® Centris Pumps as part of an innovative atmospheric water vapor isotope analyzer. Designed for fully automated operation in remote locations, these highly sensitive instruments provide environmental scientists with valuable data to help understand the shifting global climate. The National Oceanic and Atmospheric Administration (NOAA) Observatory at Mauna Loa, Hawaii
Isotopic analysis is a vital tool for climate modeling, providing important information on the movement and cycling of water and carbon around the world by measuring the ratio between environmentally significant isotopes, such as 1H and 2H or 16O and 18O. Picarro, based in Santa Clara, California, is a laboratory instrument manufacturer specializing in isotope analysis for the environmental sector, offering a range of instruments based on Cavity Ring-Down Spectroscopy (CRDS). Iain Green, Director of Business Development at Picarro, explained: “CRDS is an extremely sensitive
method which makes time-based optical measurements, thereby having higher sensitivity and precision compared to traditional absorption-based spectroscopic techniques. This makes it ideally suited to environmental isotope analysis, providing reliable, real-time data on location, without the need for significant sample preparation.” “One of the hallmarks of environmental analysis is that a set of measurements can only be performed once – the same weather pattern will never occur again – and so it is important to collect as much data as possible in real time. Our customers rely on fully automated, standalone systems which are able to operate autonomously in remote and inaccessible locations for prolonged periods. To achieve this, it is necessary to perform regular calibration checks using
known calibration standards, and so we began developing a module that would allow remote, unattended calibration of water vapor instruments, something that had never been done before.” “Vaporization of the liquid standard requires very accurate control of flow rates and injection volumes, and we needed this precision combined with extremely reliable operation that would ensure consistent performance during days, weeks or even months of continuous operation. Tecan’s Cavro Centris Pump can provide this accuracy while coping with the demanding operational needs of the application, and is compact enough to be easily integrated into our platform. The system has two Centris units – each delivering a separate calibration standard – allowing our instruments to perform optimally for prolonged periods in harsh and challenging conditions.” To find out more about Tecan’s Cavro Centris Pump, visit www.tecan.com/components
Atmospheric scientist Dr David Noone, University of Colorado, on site with the Picarro
Picarro system on location in China
To learn more about Picarro’s CRDS technology, go to www.picarro.com
MICROARRAYS TECAN JOURNAL 3/2011
Automating advances in medical technology Scientists at the Biodesign Institute Center for Innovations in Medicine have increased their sample throughput and improved inter-assay variation using Tecan’s HS 4800™ Pro microarray hybridization station. Researchers at the Center for Innovations in Medicine, part of the Biodesign Institute at Arizona State University, US, are focusing on the development of new technologies to help improve human health through advances in medical diagnostics and the prevention and treatment of disease. Postdoctoral researcher Bart Legutki explained: “I am primarily interested in infectious diseases and the early detection of breast cancer. We are using peptide microarray techniques to study proteomics, and have an extensive customsynthesized peptide library consisting of random sequence peptides. This provides an even distribution across all the possible sequences available, with a very unbiased mix. These peptides have no similarities to mammalian, bacterial or viral proteins, but do show interesting mimotope-like properties. We use a commercial grade microarray printer that allows us to print hundreds of microarrays per week to screen patient samples. As we began to investigate larger patient populations we found that manual processing introduced too much inter-assay variation and was time consuming, and so we looked for a more high throughput system that would alleviate these problems.” “We needed a system capable of handling anything from 12 to 100 arrays a day. At the time, most systems could only cope with four to eight arrays per run. We evaluated the HS Pro in our laboratory for a month using well-characterized human sera, obtaining very reproducible data, and we really liked the layout and operation. We purchased our main system in November 2009. It is now in routine use, and we plan to add another module fairly soon to increase our throughput even more.” Bart concluded: “We currently perform about 24 arrays a day – but could run 36 with shorter protocols – and we aim to increase our
The HS Pro in use at the Biodesign Institute Center for Innovations in Medicine
throughput to around 100 arrays a day soon. The HS Pro has eliminated operator-to-operator variability – allowing us to have multiple users working on our samples with one person starting a run and another person finishing it – and generates better data overall. Our scientists now have more time to perform other duties, such as data analysis, which we think will be very beneficial as we perform larger studies with more samples. The HS Pro is certainly helping to increase our productivity, and we were recently able to open our Peptide Array Core. Without the HS Pro we wouldn’t have nearly enough capacity to process so many samples.” To find out more on Tecan’s HS 4800 Pro, visit www.tecan.com/hs4800 To find out more about the Center for Innovations in Medicine, visit www.biodesign.asu.edu/research/researchcenters/innovations-in-medicine Photos courtesy of Joseph Caspermeyer
Readers TECAN JOURNAL 3/2011
Researchers at Nagasaki University develop method for antemortem CJD detection Scientists at Nagasaki University have established a sensitive diagnostic assay using Tecanâ€™s InfiniteÂŽ 200 series plate readers to detect prions in human spinal fluid samples of patients with suspected Creutzfeld-Jakob disease (CJD), offering hope for a reliable antemortem evaluation.
Dr Atarashi with the Infinite 200 PRO plate readers
Readers TECAN JOURNAL 3/2011
The Division of Cellular and Molecular Biology at the Nagasaki University Graduate School of Biomedical Sciences in Kyushu, Japan, hosts a team of researchers studying the biology of prions and their nature as infectious agents of transmissible spongiform encephalopathies (TSEs). Prion diseases are lethal neurodegenerative disorders, the most notable examples being Creutzfeldt-Jakob disease (CJD) in humans, and bovine spongiform encephalopathy (BSE), or mad cow disease, in cattle. Dr Ryuichiro Atarashi, Assistant Professor at the Division of Cellular and Molecular Biology, explained: “Prions are proteins that have the same amino acid sequence as those in healthy animals and humans but with an aberrant structure. The normal protein structure, referred to as ‘cellular prion protein’ or PrPC, is largely monomeric whereas the structure of the aberrant prion protein (PrPSc) is rich in beta-sheets and forms amyloidlike polymers. The change in structure from normal to aberrant type is induced by contact of PrPSc with PrPC, inducing aggregation.” The most common prion disease in humans, sporadic CJD, is thought to be caused by a spontaneously occurring change of the normal-type to the aberrant-type prion
protein, which then spreads and results in disease. Sporadic CJD is very rare, occurring annually in about one in a million people. In addition, heritable prion diseases, thought to make up around one or two tenths of CJD cases, are due to changes in the gene coding for PrPC and cause the patient to develop disease as they age. Together with variant CJD, which results from the transmission of prions from BSE-infected cattle, and infectious CJD resulting from transplantation of CJD-infected dura, TSEs have a significant effect on public health. Dr Atarashi said: “It has been shown in numerous animal experiments that early treatment of TSEs is critical. However, early diagnostic methods in pre-symptomatic and early stages of the diseases have not been established. In prion diseases such as CJD, the detection of PrPSc is considered a definitive diagnosis, but antemortem diagnosis requires a high-risk brain biopsy. For this reason, we have focused our research activities on developing an assay for early definitive diagnosis of CJD that allows in vitro amplification of the miniscule amounts of PrPSc contained in patients’ spinal fluid. Using Tecan’s Infinite 200 and 200 PRO series plate readers, we have been able to successfully develop just such an assay. The
method, called Real-Time Quaking-Induced Conversion (Real-Time QUIC), involves amplifying the minute amounts of PrPSc, using recombinant PrP (rPrP) expressed in E. coli as the reaction substrate. The reaction is stimulated by vigorous shaking which induces rPrP to aggregate and form fibrils. PrPSc is required for the fibril formation reaction of the rPrP; if the sample doesn’t contain PrPSc, this reaction doesn’t happen. Of course there is the possibility of a spontaneous reaction, but this would be readily distinguishable compared to when PrPSc is present.” Thioflavin T, which specifically binds to amyloid fibrils, is used to monitor the kinetics of fibril formation, with fluorescence intensity measurements taken every 10 minutes for 48 to 72 hours. Dr Atarashi added: “In testing, Real-Time QUIC has achieved greater than 80 % sensitivity and absolute specificity for the detection of abnormal-type PrP in spinal fluid from CJD patients. This demonstrates its ability for rapid diagnosis of suspected cases of CJD while the patient is still alive, offering the possibility for early treatment. Real-Time QUIC should also work well on variant CJD, although it has been difficult to obtain samples to test, and testing on heritable CJD samples is currently underway.” “Our Infinite plate readers were essential in developing the Real-Time QUIC method. It was vital to have a programmable and vigorous shaking function because we found that the more vigorous the shaking, the better the reaction. Before the introduction of the automated plate readers, we used a dedicated shaker, and prion detection was carried out laboriously by Western blots. The Infinite readers provide immediate results, measuring up to 96 samples at a time, and the process is far safer because the potentially infectious prions are sealed within the instrument.” Dr Atarashi concluded: “We have both monochromator- and filter-based models of the Infinite, and have recently introduced our fourth unit in our laboratory. We are also keen to use these plate readers for other research applications in the future.” To find out more on Tecan’s Infinite 200 PRO series plate readers, visit www.tecan.com/infinite200
Dr Atarashi and his team at Nagasaki University
components TECAN JOURNAL 3/2011
Relying on automation – new applications for circular dichroism
Automating circular dichroism measurements with the Chirascan-plus Circular Dichroism Spectrometer and the Tecan Cavro Omni Robot
Applied Photophysics has created a new protein discovery solution, incorporating Tecan’s Cavro® Omni Robot to automate sample preparation for its Chirascan™-plus CD Spectrometer. This newly created system combines Tecan’s precise and reliable sample handling with measurement reproducibility, opening up a wide range of circular dichroism applications for the first time.
The R&D team at Applied Photophysics. From left to right: Lindsay Cole (Senior Application Scientist), Paul Hatton (Senior Mechanical Engineer), Bernard Costello (Application Scientist), Rene Ayina (Senior Programmer) and Tim Collins (R&D Manager)
For over 40 years, Applied Photophysics, based in Surrey, UK, has designed and supported innovative, high performance spectrometers. The Company’s Chirascan family of instruments, which uses next generation circular dichroism (CD) technology, has opened up new areas of interest, including clone selection and biopharmaceutical formulation. Lindsay Cole, Project Leader at Applied Photophysics, explained: “CD is extremely sensitive to the conformation of biological macromolecules in solution, including secondary and tertiary protein structure, allowing us to detect significant changes in the spectrometric properties of proteins. The sensitivity of this technique requires extremely careful sample preparation to avoid reproducibility issues and erroneous results. As a result, throughput is very low when performed manually, with a study of 20 samples taking almost an entire day.” In early 2009, Applied Photophysics collaborated with the biopharmaceutical community to investigate the possibility of automating CD measurements with an OEM robot to revolutionize throughput
and reproducibility. “My project brief was to develop a system enabling unattended running of samples,” Lindsay continued. “Although we could have developed a solution in house, it quickly became apparent that the most efficient way of achieving reliable automation was to use an existing system from an OEM supplier. We looked at the various systems on the market, and it was obvious that the Tecan Cavro Omni Robot best suited our needs. There are various ‘HPLC-type’ autosampling systems on the market, but these would have significantly constrained our instrument design. In comparison, the Omni Robot is a gantry format general-purpose component that can simply be bolted onto our platform, offering the required xyz liquid handling without further modification. This eliminated the need to design and integrate our own fluidics, sample probe and injection port, and gave us useful features, such as the sample level sensing, and the option to add other sample handling and preparation capabilities in the future. Tecan engineers also came to our facility and integrated an Omni Robot with our Chirascan-plus instrument as a proof-ofconcept – which worked first time – and so
components TECAN JOURNAL 3/2011
we were confident that the Tecan solution offered exactly what we needed.” “Because the Tecan component is a relatively open and malleable liquid handling robot, it was very easy to incorporate it into our system. The Omni Robot is held in a purpose-built enclosure alongside the Chirascan-plus, and we had a basic system set up and working within a month of receiving our first unit. All our efforts could then be focused on designing the critical fluidics and sample chamber to suit an automated set-up, as well as developing experimental sampling and cell cleaning protocols.” The automated platform uses a single fixed probe and a Cavro XCalibur syringe pump to perform sequential sampling from up to four 96-well plates, without dilution or cross-contamination of samples. Auxiliary components – such as the sample chamber lid, liquid wash pumps and vacuum pump – are controlled by a Cavro Smart I/O Board in the Omni Robot. The system is also able to automatically switch between dispensing water, cleaning solution or solvent via a smart
distribution valve, allowing thorough cleaning and drying of the sample cell between measurements. “Sampling, measurement and cell cleaning take up to ten minutes per sample, and we can run up to 200 samples per day, or 384 samples over a weekend, with just about 30 minutes of plate loading and experimental set-up by a technician. The instrument is temperature controlled to avoid sample degradation during extended runs, and we can also use the Omni Robot to prepare titration plates with variations in pH, salts and ligands, to allow easy investigation of conditions that destabilize the conformation of target macromolecules,” Lindsay added. “Automation significantly increases the practical throughput of the Chirascan-plus, allowing unattended running with minimal
Increasing throughput with the Tecan Cavro Omni Robot
user intervention. It has reduced labor requirements per measurement by 90 %, with a 10-fold increase in productivity, so in terms of samples per person per hour, it increases productivity about 100 times. This, combined with the reliability and increased reproducibility offered by the Cavro Omni Robot, has greatly increased the appeal of CD as an analytical technique for proteins, particularly for the biopharmaceuticals development market.” To find out more about Tecan’s Cavro Omni Robot, visit www.tecan.com/components For more information on Applied Photophysics, visit www.photophysics.com Chirascan is a trademark of Applied Photophysics.
LIQUID HANDLING & ROBOTICS TECAN JOURNAL 3/2011
Pressure Monitored Pipetting supports Swiss BioQuant’s dedication to excellence The Pressure Monitored Pipetting (PMP™) option for Freedom EVO® workstations is helping to ensure reliable sample preparation at Swiss BioQuant, allowing real-time quality control while pipetting plasma, urine and other complex matrices, and supporting the Company’s dedication to service excellence.
Stefan Schipp (left) and Christoph Siethoff of Swiss BioQuant
Swiss BioQuant is a bioanalytical contract research organization (CRO) based in Reinach, near Basel, Switzerland. The Company specializes in LC-MS/MS, performing customized method development, validation and bioanalysis for biotechnology companies and pharmaceutical manufacturers. Stefan Schipp, Staff Scientist at Swiss BioQuant, explained the Company’s activities: “We receive both plasma and urine samples from a wide range of animal model studies and clinical trials, using mass spectrometry-based analytical techniques to identify and quantify molecules of interest to our customers. Samples arrive in our laboratory directly from these studies, and are prepared for LC-MS/MS analysis in house using protein precipitation and solid phase extraction methods.”
As a CRO, reliable and reproducible sample handling is vital. Automation is the most effective way of achieving this, and the Company purchased a Freedom EVO 100 workstation specifically to automate sample preparation for the majority of clinical study work. Christoph Siethoff, President and CEO of Swiss BioQuant, said: “At my former workplace we had three systems from Tecan; we had experience of the liquid handling workstations and knew the flexibility of the Freedom EVOware® software and, in our experience, Tecan systems are robust and reliable. However, we deal with difficult matrices every day and pipetting of both plasma and urine samples can be challenging, as clogging of pipette tips – due to clots and other aggregates within the sample – is a common problem. The option of a PMP module to deal with this problem confirmed our decision to choose Tecan’s equipment. The PMP option checks all our liquid handling processes, allowing real-time monitoring of critical aspiration and dispensing steps to identify any pipetting errors. Without this option, we could never be absolutely sure that we were pipetting the correct volumes; our only option was to try to check the liquid levels visually, but that’s neither easy nor reliable in a 96-well plate. PMP is easy to use and ensures that any errors are immediately identified, giving us the option to rectify the problem or repeat samples as necessary. We don’t need to continuously watch the system and we’re no longer running the risk of generating erroneous results.”
As well as the PMP option, Swiss BioQuant’s platform is also equipped with both an 8-channel Liquid Handling (LiHa) arm and a Te-VacS™ module, allowing completely automated processing of samples for LC-MS/MS analysis. Stefan added: “For each study we are able to develop a specific method using Freedom EVOware, modifying the individual scripts according to the customer’s sample type and individual requirements. This flexibility in method development significantly accelerates processing, and the software enables full sample traceability throughout preparation.” Christoph concluded: “PMP very much supports our Company’s dedication to service excellence and is an important asset in discussions with our customers. In the past, some customers have questioned whether manual sample preparation procedures are better than automation, but PMP gives us far better sample security than we could ever achieve in manual processes. Our customers can be sure they are getting reliable results and we are confident that we are providing them with a first-rate service.” To find out more on Tecan’s Freedom EVO liquid handling platforms, visit www.tecan.com/freedomevo To learn more about Swiss BioQuant, go to www.swissbioquant.com
LIQUID HANDLING & ROBOTICS TECAN JOURNAL 3/2011
Pressure monitored aspiration being performed on the Freedom EVO
READERS TECAN JOURNAL 3/2011
Sunrise™ in Mongolia
Scientists involved in the study of vitamin D levels in the Mongolian population were in need of a microplate reader to help further their research. An amazing collaboration evolved, with staff from Tecan, CERLab, and Brigham and Women’s Hospital working together to solve the problem.
Mongolian schoolchildren in class
Mongolians are at high risk of vitamin D deficiency because they reside at high northern latitude where, although it is always sunny, low temperatures prevent exposure of the skin for most of the year. In addition, the sun hits the earth at such an oblique angle that the UVB radiation – which is required for natural vitamin D synthesis – has to travel through a thick ozone layer that filters it until it is very weak, further limiting production of vitamin D. This situation is compounded by not eating fish and a lack of availability of vitamin D-fortified foods. Scientists at the Health Sciences University of Mongolia have been investigating the issue, as Dr Janet Rich-Edwards, an Associate Professor at Brigham and Women’s Hospital, Harvard Medical School, Massachusetts, US, explained: “We have been working in Mongolia since 2004, and have seen the incredibly high incidence of rickets in the country. Vitamin D deficiency was considered an infant problem, because its most obvious symptom is rickets. In fact, vitamin D deficiency affects the entire population. Rickets was thought to be the tip of the iceberg; it was suspected that vitamin D deficiency was widespread across Mongolia, and yet there was no readily available vitamin D assay.” “Our original vitamin D pilot study involved a small group of children. At that stage there was no way of performing the assay in Mongolia, and we were forced to transport frozen samples to the US for testing, which was a logistical nightmare and a huge barrier to research. Gary Bradwin, Manager at the Clinical and Epidemiologic Laboratory (CERLab), Children’s Hospital Boston, became involved, volunteering his time to help Mongolian scientists set up the assay and perform the vitamin D analysis themselves.”
Gary added: “CERLab is a reference laboratory offering clinical chemistry services to laboratories worldwide, and we became involved with the project during the initial pilot study. I visited Mongolia to help set up the assay and we looked at all the different methods available, eventually opting for ELISA as the cost per sample is not too high and all that was really needed was a plate reader. However, the laboratory was very short of equipment and consumables and it was heartbreaking to see just how poor they were. Consumables that we take for granted as disposable, such as pipette tips and needles, were sterilized and reused. Tests also had to be performed on serum as there was no centrifuge to separate whole blood; instead they had to wait for the blood to clot and then remove the serum. The plate reader was ancient and could only handle one eight-well strip at a time, so they needed an assay that could break down into removable strips, which not all ELISAs are able to do. Performing an ELISA like this is very slow and you need to be extremely careful; it is so much easier with instruments such as the Sunrise, which can read an entire plate in just a few seconds. Data handling was also problematic; it was not computerized, relying instead on a small printer that simply recorded the raw absorbance numbers. To analyze actual samples, they had to draw the calibration curve manually on graph paper and then read the samples against it.” “The experience made me appreciate everything my laboratory had, and I really wanted to try to help make life easier for the Mongolian investigators. We had a Sunrise reader that we were not using, so I decided to make sure it was operational and then donate it to the Mongolian laboratory.
READERS TECAN JOURNAL 3/2011
Initially, I had difficulty getting the Sunrise reader to communicate with the Magellan™ software and so I contacted Tecan.” Leith Olson, Technical Service Specialist at Tecan, takes up the story: “Gary contacted our Helpdesk in October 2010 because he couldn’t connect to his Sunrise reader with Magellan. While we were attempting to troubleshoot the instrument, he mentioned that he wanted to get it up and running again so that he could donate it to a laboratory in Mongolia that was trying to establish a program to measure vitamin D in the country’s children. I realized that the Sunrise would need to come in for servicing, which we did free of charge in support of this worthy cause. We also wanted to perform a firmware upgrade, and so the system was sent to Tecan Austria for refurbishing.”
Gary continued: “I was amazed to discover that my Sunrise had been sent to Austria, and I really could not believe how much work Tecan had done to rebuild the system. It was completely unrecognizable; almost everything seemed to have been replaced, even the case. When the Mongolian investigators received the Sunrise, they were overwhelmed. The reader is easy to use and, as I gave them all the manuals, they were able to set it up themselves. The computerized data handling has made life so much easier for the scientists; they couldn’t quite believe that it was possible to just read a plate and automatically generate the results, and it’s so much faster than their previous system. The Sunrise is working beautifully and has totally transformed their workflow.”
Mongolian children undergoing a medical check-up during the study
Janet concluded: “The Sunrise is serving as a real fulcrum in enabling the first national survey of 25-hydroxyvitamin D levels, and this will directly contribute to setting Mongolian national policy on food fortification and the possible eradication of rickets.” Gary added: “I was really impressed that Tecan took so much interest and went as far as they did. They’ve certainly made a fan here.” To find out more on Tecan’s Sunrise microplate reader, visit www.tecan.com/sunrise To find out more on Harvard Medical School, visit www.hms.harvard.edu/hms/home.asp To find out more on Brigham and Women’s Hospital, visit www.brighamandwomens.org/
Phlebotomist taking a blood sample from a Mongolian child
Agriculture TECAN JOURNAL 3/2011
Freedom EVO® in Evogene’s Biolab runs 13 cloning related procedures Automation with the Freedom EVO system and SciRobotics’ Pickolo™ colony picker has revolutionized the gene cloning and colony picking workflow at Evogene’s laboratory, allowing timeefficient and reliable tracking of cloning processes and enabling scientists to increase the yearly production of clones.
Evogene is a world leader in developing improved plants for the agriculture and biofuel industries through the use of plant genomics. Based in Rehovot, Israel, the Company provides a complete solution for improving plant traits, such as yield or drought tolerance, for several agricultural companies, using advanced breeding methods and its own proprietary computational genomic technology. The process of discovering and targeting new plant traits combines numerous technologies, including high throughput plant validation systems, field experiments and genomic data creation.
that are then passed over to the Plant Transformation Unit for transformation into a model plant. Many of the steps involved at this stage, such as PCR amplification, DNA restriction and ligation, clone screening and sequencing, require specific clone design and a different set of restriction enzymes, as well as different sets of unique primers. All of these procedures need to be handled with the highest attention to detail; the success of every step in the process needs to be tracked and all the intermediate products need validation. Performed manually, this process is time-consuming, labor-intensive and error-prone.
As part of the process, the Biolab team needs to clone and amplify several hundreds of genes of interest in a limited time span
Dr Alin Sela-Brown, Head of Evogene’s Biolab, explained the challenges of the manual cloning process in her lab: “The colony PCR screening days used to be a very stressful time in our laboratory. Hundreds of colonies had to be picked into hundreds of PCR reactions, each composed of a different primer set.” On the same day, the colony PCR product was loaded onto a gel for electrophoresis and two positive colonies from each clone were chosen. These were then selected and inoculated into overnight cultures for plasmid purification. “To achieve this we had to ‘borrow’ many laboratory members performing other projects and assignments to help us with this PCR screening task and the process was very intensive.” “Due to the large amount of genes that we have to clone, we knew that a standalone picker performing only a single task would not be enough and realized that, by integrating colony picking into the liquid handling platform, Tecan offered us exactly the throughput we needed.”
Dr Alin Sela-Brown (Evogene, front) and Dr Shai Kaplan (Head of R&D, SciRobotics Ltd) with the Freedom EVO workstation
Agriculture TECAN JOURNAL 3/2011 The deck layout of Evogene’s Freedom EVO platform
Primer normalization and stock solution
Frozen stock preparation
Colony screening PCR set-up
Culture inoculation Sequencing preparation
Heat shock transformation
Restriction and ligation
The Freedom EVO 150 system was configured with a Liquid Handling arm with eight channels, a Te-Shake™ shaker, a Te-VacS™ vacuum separation module and SciRobotics’ integrated Pickolo colony picker. The system performs several tasks for the Biolab, including colony picking, primer normalization and stock dilution, PCR amplification and DNA purification, restrictions and ligations, agar plate dispensing and preparation of frozen stocks.
• Primer normalization and stock dilution – The instrument automatically normalizes the primer and dilutes for working stock according to the primers order file received with the primer plate.
“Colony picking was our main process bottleneck but we also wanted to automate other main upstream and downstream applications – PCR set-up and DNA extraction – on the instrument. We were very satisfied when our Freedom EVO was able to run not just these three processes but, in fact, thirteen of them!”
• Ligation – The restricted amplicon of each clone is ligated to a suitable plasmid.
“Before we had the Freedom EVO system, colony picking and PCR screening required very intense and focused days. Many people from our lab were recruited and devoted only to this task. Now, it takes just one person a few hours, leaving enough time for downstream processing and parallel activities. And the stress-free process we have with our Freedom EVO system drastically reduced the errors.”
• PCR amplification – The instrument matches the correct template with the correct primers according to the database information. • PCR purification – PCR purification is performed using the Te-VacS, with purification products tracked automatically. • Restriction – The amplicon is restricted with the clone-specific restriction enzymes.
• Colony screening – A unique primer is used for each clone to allow screening. • Colony picking – The Pickolo automatically identifies positive colonies.
The ability to handle any subset of clones and perform time-consuming procedures on the Freedom EVO system has provided Evogene’s Biolab with the flexibility and speed it needed to respond to its growing requirements. The integration of the Freedom EVO workstation and SciRobotics’ Pickolo colony picker saves time both at the bench and in experimental planning, creating an efficient and reliable cloning workflow.
Agar plate preparation
• Culture inoculation – Positive colonies are inoculated to starter cultures and solid agar stock replicates and incubated overnight. • Freeze-stock preparation – Glycerol freezestocks of positive colonies are prepared. • Plasmid miniprep – The instrument performs plasmid purification using the Te-VacS with a specific kit. • Sequencing preparation – The instrument prepares several sequencing reactions for each clone with its own sequencing primers and automatically prepares documentation of the sequencing plate. • Heat shock transformation – The instrument performs the necessary pipetting steps of the heat shock procedure. • Agar plate preparation – The instrument dispenses the agar media from a stirrer into 96-well plates.
To find out more on Tecan’s Freedom EVO liquid handling platforms, visit www.tecan.com/freedomevo To learn more about SciRobotics, go to www.scirobotics.com To learn more about Evogene, go to www.evogene.com Pickolo is a trademark of SciRobotics.
Forensics TECAN JOURNAL 3/2011
Automation of DNA analysis boosts the fight against crime The DNA Analysis group at the Hessen Landeskriminalamt in Germany has automated DNA trace analysis with the HID EVOlution™ System, increasing throughput of crime samples, providing results in approximately three days and improving the quality of analysis.
The DNA Analysis group at the Hessen Landeskriminalamt, Germany
The Landeskriminalamt (LKA) in Wiesbaden is the main headquarters of the criminal investigation department in the German State of Hessen. The LKA includes a Department of Forensic Science that serves all police departments, prosecutors and courts in the district, and, within this, a group dedicated entirely to examining evidence that carries organic traces or textile fibres. Dr Diane Schmidt, leader of the DNA Analysis subgroup investigating burglary and theft, described the laboratory’s workload: “Last year, we analyzed 50,000 traces received from around 4,000 criminal cases, collected from all kinds of objects, from furniture to bottles, tool kits, door handles, simply everything; everything people touch or work with can carry DNA traces. At the time we analyzed 20,000 of these with the HID EVOlution System, and a proportion was outsourced to local forensic medical institutes. In the long term, however, we had been looking at how automation could help us to bring as much of this work in house as possible, to give us the capacity to handle the anticipated increase in cases and to give us tighter control on our samples and their analysis.” Diane was involved in the initial choice of an automated system in 2007 after looking at several available systems and sharing the experiences of other state forensic departments in Germany. “Several factors influenced our decision to choose the HID EVOlution System,” she said. “Our colleagues in other LKAs had positive experiences using the system, and the system is very flexible, handling sample preparation from different sources if required. The combination of the reliable Freedom EVO®
platform and dedicated software from Tecan with scripts developed by Applied Biosystems (Life Technologies) was very important to us, and produced a really useful prevalidated system for forensics. Although we still needed to perform our own validation within the laboratory to adjust the system to our particular requirements, we could rely on many things that were part of the pre-validation package, and this certainly made the whole process much easier. The HID EVOlution, together with the Applied Biosystems PCR systems and genetic analyzers, is able to address the handling of all five steps of trace analysis – extraction, quantification, amplification, genetic analysis and evaluation of results – but it is very user-friendly and easily managed by our six technical assistants. For crimes such as theft or burglary, the samples are put directly onto the ‘street’ (the laboratory workflow) on the HID EVOlution System and we will have the results within three days.” The first phase is DNA extraction using the HID EVOlution extraction system and the PrepFiler® forensic DNA extraction kit, followed by amplification by real-time PCR, and quantification on the Applied Biosystems 7500 Fast Real-Time PCR System. The next step is simultaneous amplification of this DNA with eight markers required by the German DNA database and three additional STR loci required by the European Network of Forensic Science Institutes (ENFSI). On the next day, amplified DNA samples are genetically analyzed – with 80 samples per cycle – in the Applied Biosystems 3130xl Genetic Analyzer, and results are evaluated with GeneMapper® ID-X software.
Forensics TECAN JOURNAL 3/2011
The HID EVOlution extraction, HID EVOlution qPCR/STR and Freedom EVO CE set-up systems
“The purity of the DNA samples extracted on the HID EVOlution System is excellent,” Diane added. “The extraction chemistry handles our range of different trace materials very well, and the system itself is very reliable. The HID EVOlution workstation extracts comparable quantities of DNA to our manual extraction method (see Table 1), and we are able to achieve higher levels of PCR amplification from the same amount of starting DNA, which is important when the quality of DNA can sometimes be poor due to degraded samples. We still manually analyze traces from serious crimes, such as murders, because results are required by the next day, but automation really comes into its own when analyzing large numbers of samples. The DNA ‘street’ automated laboratory workflow has helped us to achieve extremely short processing times of just eight to ten weeks for burglary trace analyses – a processing time unique throughout Germany – and another big advantage is the collection of data not only from single cases but also on mass crime in the area.”
The laboratory in Wiesbaden currently handles trace analysis for both minor and serious crimes from two police headquarters in Hessen, and only serious crimes for the remaining police headquarters, with their traces from minor crimes handled by the local forensic medical institutes. An additional HID EVOlution System is in the pipeline, taking advantage of the efficient automation process that has been developed, and the Department is confident that it will soon be able to handle the anticipated increase in workload to 60,000 trace samples a year. Diane concluded: “We are very happy with the system and have
had great support from Tecan and Applied Biosystems (Life Technologies) in setting up the right solutions for our laboratory.” To find out more about the HID EVOlution System, visit www.tecan.com/hid For more information on the Hessen State Criminal Office, visit www.polizei.hessen.de/hlka/ PrepFiler and GeneMapper are registered trademarks of Life Technologies, Inc.
Table 1: Comparison of DNA quantity extracted from blood discs with the standard manual method and the HID EVOlution System Number of blood discs
Average DNA yield [ng/μl] Manual method
Comparative yield (% of manual yield)
Comparison of DNA extracted from blood discs with the standard manual method and the HID EVOlution System
ENVIRONMENT TECAN JOURNAL 3/2011
Second generation biofuels – making energy from waste
The Centre for Novel Agricultural Products (CNAP), a specialized research center within the Biology Department at the University of York, UK, is using Freedom EVO® workstations in its research into the potential of plant-and microbial-based renewable resources. Left to right: Simon McQueen-Mason, Caragh Whitehead, Leonardo Gomez and Poppy Marriott
Plant biomass is one of the greatest reserves of fixed carbon on the planet and, as such, has huge potential as a renewable energy source to replace fossil fuels. The main structural material in plants, lignocellulose, is composed of cellulose, hemicellulose, and lignin and has a long history as a source of energy. A drawback, however, is that the carbon required is largely present in cell walls, and is not easily accessible. A 20-strong group based at the University of York and led by Professor Simon McQueen-Mason is looking closely at plant cell wall biology, and investigating ways of successfully and cost-effectively releasing and processing the lignocellulose biomass from cell walls. In contrast to first generation biofuels which are produced from food crops, the York Group is concentrating on saccharification as a method of breaking this complex carbohydrate into its monosaccharide components, opening doors into a second generation of biofuels potentially produced from food crop waste such as leaves and stems.
In order to screen thousands of plant samples, both from classical breeding experiments as well as specifically created plant mutants and different enzymes, the Group needed to develop ways of miniaturizing and automating the saccharification assay and turned to Tecan for a solution. Dr Leonardo Gomez, who developed the assay, said: “We approached Tecan because of its reputation for developing flexible, reliable and easy-to-use automated systems, and we have certainly found that to be true. We first chose a Freedom EVO 200 workstation that is very flexible and we have used it for other applications besides the standard saccharification analysis, including the determination of the kinetics of enzyme preparations, the activity of cellulases and in comparing a matrix of different enzymes and substrates.”
In the automated process, ground and aliquoted plant samples of a defined particle size first undergo a mild pre-treatment by heating the samples in the presence of alkaline or acid solution, followed by hydrolysis and finally a colorimetric detection assay for the free sugars based on MTBH (3-methyl-2benzothiazolinonehydrozone). Pre-treatment, hydrolysis and the assay steps of sugar determination are all performed on the Freedom EVO system, equipped with a Liquid Handling arm with eight fixed tips, a sample loading area, a heated shaker, a Monitored Incubator Option (MIO™) shaking incubator and three thermocyclers. The thermocyclers achieve even and fast heat distribution across the microplates which has proven to be crucial to the MBTH assay. Figure 1 shows an overview of the saccharification process and the employed hardware and labware.
ENVIRONMENT TECAN JOURNAL 3/2011
Saccharification assays of plant material are prone to variation due to the heterogeneous and largely insoluble plant samples and the use of crude enzyme mixtures. Automation of the assay removes this problem and has improved the overall assay by standardizing conditions, like incubation timings. This was clearly seen in a comparison of the manual method with the automated protocol which showed a decrease in the coefficient of variation from 9 % to 5.5 %, dramatically increasing the repeatability and comparability of the test results. Depending on the conditions, 80 samples in quadruplicates on
four microplates are typically analyzed per day on York’s first platform. However, as the interest in second generation biofuels grew, the Group acquired a second Freedom EVO system to increase the number of analyses it can perform in its laboratory. Leo concluded: “We are very pleased with our choice of automated platform; we have successfully miniaturized this process and created an efficient and reliable workstation that handles high throughputs with ease.”
See the Group’s Freedom EVO workstation in action and described clearly in the video entitled ‘High throughput saccharification assay for lignocellulosic materials’ at www.jove.com/details.php?id=3240 To find out more about Tecan’s Freedom EVO liquid handling platform visit www.tecan.com/freedomevo To find out more about the Centre for Novel Agricultural Products (CNAP), visit www.york.ac.uk/org/cnap/
Figure 1: Labware, hardware and incubation conditions used in the saccharification analysis Process
standard 1 standard 2 standard 3 standard 4 sample sample
Determination in triplicate
Hardware Heating block
QUALITY ASSURANCE TECAN JOURNAL 3/2011
Quality matters: audits as a learning platform Quality audits are a requirement of ISO certification, and are performed to verify that standards are being conformed to by reviewing objective evidence. Tecan has introduced an internal training program for quality auditors, helping to maintain the Company’s high quality standards and customer satisfaction.
The word audit comes from the Latin audiō, which means ‘to hear or listen’. An audit is a useful, hands-on management tool which helps companies and organizations to continuously improve their performance and achieve greater customer satisfaction. The international standard for auditing quality and environmental management systems within a company is clearly defined in ISO 19011: 2002, providing a clear explanation of the guiding principles for management of audit programs and how to plan and conduct an audit, either internally or externally, as well as the skills, competencies and behaviours required for auditors. In addition, various government organizations – including the FDA (US), ANVISA (Brazil) and CMDCAS
Giuseppe Grignano and Giuseppe Simonetta
(Canada) – require additional validation that ensures quality management systems are effective, adequately implemented and sufficiently documented. Within Tecan, auditing is used as a vital tool to strengthen the Company’s values and to create solid foundations from which to achieve its vision. Giuseppe Grignano, Global Compliance Validation Manager, explained: “To ensure the future success and security of our business, it is vital that we maintain and enhance our quality standards, and auditing is an important management tool for assessing our performance in this area. Learning is a lifetime exercise for both people and organizations and, for maximum benefit,
an audit should not only report on areas of concern and non-conformances, but also highlight areas of good practice, allowing us to learn from each other to improve our working practices and enhance our business.” “Companies usually send staff on external training courses to gain the required expertise and accreditation for quality auditing, but we have initiated an internal training program to maintain and continually improve the expertise of our auditors. Working with Bureau Veritas – a world leading independent company in conformity assessment and certification services – the Tecan Operational Excellence Team developed a course that explains the audit
TALK TO TECAN TECAN JOURNAL 3/2011
process and its benefits. This initiative brings together over 40 auditors from various areas of the business, including research and development, manufacturing engineering, procurement and quality, helping to expand and strengthen the skills of both new and existing auditors.” Run at sites in Switzerland, Austria and the USA, the two-day course uses a combination of classroom-based presentations and role playing exercises. Giuseppe continued: “Participants learn about the requirements of ISO 19011, the behavior expected by auditors when conducting audits and how to resolve the needs of conflicting interests through constructive discussion. At the end of the course there is an exam based on the ISO 19011 standard. This includes both theoretical situations and practical examples of current challenges, often yielding solutions that can subsequently be applied to the business. Exam papers are sent to Bureau Veritas for external verification and, on successful completion, Bureau Veritas awards a certificate of accreditation to participants.” Giuseppe Simonetta, Global Quality Manager, concluded: “As a company, we are continually working to improve our expertise in this area, improving the skills of our auditors and enabling them to act as leaders in their core competencies. This ensures we are able to fulfil the regulatory compliance needs of the market and, ultimately, secure the future of our business. This initiative will help us to improve operational efficiency and give our customers complete confidence in our auditing processes, because our auditors have been examined and approved, not by Tecan, but by an external, independent company.”
Gerald Schmidt, Head of Sales – Europe, Near East, Middle East and Africa
Leading the Debate Since moving to Tecan six months ago, I have soon come to realise that this Company thrives on an inspiring and stimulating business culture. From its strong base in Europe, Tecan is a truly global company in its thinking, working effectively at a local level and maintaining essential links with teams in the US and Asia Pacific. At the core of Tecan’s vision is quality and reliability on a global scale; it is clearly a major life science business and has the competence, understanding and innovation to exceed the expectations of our customers in life science laboratories around the world. We are not just saying this, we really can do it. Tecan has long been renowned for its engineering capabilities, however, I had completely underestimated the Company’s real strength in life sciences. Within Tecan there is a huge breadth of international knowledge in all life science fields, from genomics to proteomics, cell biology to metabolomics, and most of the product management and application specialists are life science- not just engineering-based. This knowledge is used to transform time-consuming manual processes into state-of-the-art automated solutions for various ground-breaking technologies and sophisticated workflows. Applying these solutions provides a fast response to the demands of the market and that, for me, is really true innovation. Email email@example.com to tell us your experiences of Tecan’s innovation in action.
EVENTS 2011 TECAN JOURNAL 3/2011
Meet Tecan at these events from now until the end of this year Americas MicroArray World Congress 2011
South San Francisco, USA
29 - 30 Sept 2011
ICHG International Congress of Human Genetics 2011
11 - 15 Oct 2011
AABB American Association of Blood Banks 2011
San Diego, California, USA
22 - 25 Oct 2011
AAPS American Association of Pharmaceutical Scientists
Washington, DC, USA
23 - 27 Oct 2011
BPI BioProcess International 2011
Long Beach, California, USA
31 Oct - 04 Nov 2011
Discovery on Target
02 - 04 Nov 2011
ASCB American Society for Cell Biology
Denver, Colorado, USA
03 - 07 Dec 2011
70th Annual Meeting of the Japanese Cancer Association
03 - 05 Oct 2011
AACB50 Australasian Association of Clinical Biochemists Golden Jubilee Conference
10 - 14 Oct 2011
12 - 15 Oct 2011
Asia and Pacific
03 - 04 Nov 2011
MBSJ Molecular Biology Society of Japan 2011
13 - 16 Dec 2011
20 - 22 Sept 2011
11 - 13 Oct 2011
Europe, Middle East and Africa
14 - 18 Nov 2011
16 - 19 Nov 2011
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