Tecan Journal

Transcription

Tecan Journal

Tecan Journal
Edition 2 / 2007
From bluetongue to bird flu,
detecting infectious diseases
with the Freedom EVO®
page 7
High throughput
nucleofection with amaxa
technology
pages 9 – 11
New tools for predicting the
recurrence of breast cancer
pages 12 – 13
C E O WE LCOME
Welcome...
...to this, the second issue of the Tecan Journal for 2007.
Tecan Journal 2/2007
Our recently published financial results
confirm that 2006 was a year of strong
growth for our company and I would
like to personally thank you all for your
continued confidence and support.
Our successful year can, of course, be
attributed to our innovative products, but
also to our ongoing efforts to improve
and streamline our organization. Our
companies around the world share
the common goal of perfecting their
quality management systems for
all manufacturing, sales and service
activities, and continually strive to listen
more closely and respond more effectively
to feedback from you, our customers.
There are updates from customers
working in a range of diverse applications,
from avian influenza and nucleic acid
testing to forensics and cell biology, who
count on our systems to produce the
consistently reliable results they need
every day. We are also delighted to share
with you details of exciting new product
developments which demonstrate our
commitment to continually improving
and expanding the breadth of products
we offer.
In this issue, we bring you news of
several carefully-chosen collaborations
with other life science companies. Many
of these partnerships add additional
capability to Tecan’s liquid handling
workstations and increase their range
of applications, providing you with even
more application-specific automated
laboratory solutions.
Thomas Bachmann, CEO
I hope you enjoy this issue of the Tecan
Journal, and look forward to another
successful year.
CONTE N TS
CEO welcome
Thomas Bachmann, CEO, welcomes you
to this issue of the Tecan Journal.
page 2
Automated virus testing in animals
From bluetongue to bird flu, detecting
infectious diseases with the Freedom EVO®.
page 7
Corporate news
Find out about some of Tecan’s recent
collaborative projects.
page 4
Working for ISO accreditation around
the world
Implementing a new quality management
system through all Tecan’s sales and
service departments.
page 8
High throughput nucleofection
with the Freedom EVO workstation
and amaxa 96-well Shuttle®
Fully automated, high throughput
transfection of primary cells and
difficult-to-transfect cell lines.
pages 9-11
Product news:
Painting a picture of 20,000 genes
in the mouse brain
Tecan’s GenePaint™ systems are used
to create a groundbreaking new gene
expression atlas.
page 5
7 detection modes
Tecan’s highly sensitive and extremely fast
Safire2™ and Infinite™ F500 microplate
readers offer seven different detection
modes.
page 5
The target of personalized medicine
moves ever closer
Genomic Health in California develops
tools to predict breast cancer recurrence
using Tecan workstations.
pages 12-13
Genetic fingerprinting
of crime scene samples
Forensic scientists in Stuttgart, Germany,
have commissioned a semi-automated
sample processing system from Tecan.
pages 14-15
Engineering new biomaterials
for human implants
Optimizing automated tissue culture
methods with the Freedom EVO liquid
handling workstation.
pages 18-19
Plasma technology for fast,
effective tip cleaning
Tecan teams up with Cerionx to combine
‘cold’ plasma tip cleaning systems with
the Freedom EVO.
pages 20-21
Building a cattle DNA database in Japan
Automated DNA extraction for analyzing
cattle parentage with the Freedom EVO
200.
pages 22-23
Managing worldwide compound delivery
with REMP technology
Boehringer Ingelheim’s Screening Support
group has a number of REMP systems
to manage its library of more than
800,000 compounds.
pages 24-25
Cellerity™ is now available with
the MultiChannel Arm™
The MCA brings increased speed and
throughput to automated cell culture.
page 5
HS Pro: miRNA profiling
and Exiqon LNA™ arrays
Profiling the expression patterns of
hundreds of microRNAs with the HS
4800™ Pro Hybridization Station.
pages 16-17
Freedom EVOlyzer® received with great
enthusiasm in Spain
The automated ELISA platform satisfies
a wide range of clinical diagnostics
laboratories.
page 6
Te-PoolSafe™ boosts pipetting accuracy
for testing donor blood
The German Red Cross Institute screens
thousands of donated blood samples every
day and has validated Tecan’s liquid arrival
check system.
pages 26-27
Events 2007
Meet Tecan at a number of events around
the world over the next six months.
page 28
CO R PO R AT E N EWS
Tecan teams up with high profile business
partners in a number of new developments
In recent months, even more collaborations with
industry partners have come to fruition, bringing
application-specific, automated laboratory systems
to researchers working in a range of disciplines.
Tecan and Applied Biosystems have
jointly developed the HID EVOlution™
System for the automated analysis
of forensic DNA casework samples.
This new standardized system
provides fully automated sample
quantitation, normalization and short
tandem repeat (STR)-PCR set-up, by
integrating Tecan’s Freedom EVO® 150
liquid handling workstation with the
Applied Biosystems 7500 Real-Time
PCR System, 3130xl Genetic Analyzer,
and Quantifiler® and AmpFℓ STR®
DNA testing kits. The HID EVOlution
is expected to reduce the amount
of time required to process forensic
DNA samples and help laboratories to
increase their sample throughputs.
Tecan and amaxa, Inc. have successfully
integrated amaxa’s Nucleofector®
96-well Shuttle® System with the
Tecan Freedom EVO liquid handling
workstation, allowing fully automated,
high throughput transfection of
primary cells and difficult-to-transfect
cell lines, including non-dividing neurons
and T cells, with siRNA, DNA and other
substrates. The development is ideal
for large-scale studies that involve
high throughput transfection stages,
such as RNAi-based screening for drug
development. The 96-well Shuttle® relies
on unique electroporation technology for
transfection of primary cells with high
efficiency and excellent survival rates.
Integrating this system with the Freedom
EVO workstation entirely removes
all manual steps for cell preparation
and transfection, allowing 96-well
microplates to be completely processed in
just minutes, depending on the protocol
used. You can read more about the
collaboration on pages 9-11.
Tecan and VWR International, Inc. have
announced a strategic alliance to unite
the strengths of each company and
together provide Tecan’s cutting edge
microarray instruments and microplate
readers to the global research market.
The collaboration takes advantage of
VWR’s BioSciences program, which aims
to provide researchers with the highest
quality services and products available,
whatever their application, and a
comprehensive sales network – including
more than 120 sales representatives
worldwide and 15 BioSciences managers
in Europe alone. Tecan’s range of
technically excellent microarray products
and microplate readers offers perfect
solutions for customers of the BioSciences
program working in all areas of drug
discovery and life science.
Tecan and Cerionx, Inc. have made
an exclusive distribution agreement
for the supply and marketing of the
Cerionx 8-channel TipCharger™ System
to the worldwide biopharmaceutical
market. Addition of the TipCharger™
System as an integrated and supported
component to Tecan’s automated liquid
handling and robotics workstations offers
uniquely superior pipette tip cleaning
and sterilization. The TipCharger™
System uses room temperature (‘cold’)
atmospheric pressure plasma to clean
and sterilize pipette tips by removing
contaminants at the molecular level
without using hazardous solvents. This
revolutionary plasma cleaning technology
reduces the potential for contamination
at the sample and consumable levels,
reduces run time and improves assay
productivity. Find out more about the
Cerionx TipCharger™ System on pages
20-21.
Quantifiler and AmpFℓ STR are registered
trademarks of Applera Corporation or its
subsidiaries in the US and/or certain other
countries.
The Nucleofector 96-well Shuttle System is
covered by patent and/or patent-pending rights
owned by amaxa.
TipCharger is a trademark of Cerionx, Inc.
PRODuCT N EWS
Painting a picture
of 20,000 genes in
the mouse brain
Tecan’s GenePaint™ systems for
automated, high throughput in situ
hybridization have been used to develop
a groundbreaking, genome-wide atlas of
gene expression in the adult mouse brain.
The comprehensive digital atlas details
the expression patterns of approximately
20,000 genes, providing an invaluable
source of information for scientists
studying brain organization, gene
functions and behavior. Watch out for a
special feature on the project in the next
issue of the Tecan Journal!
7 detection
modes
Tecan’s highly sensitive and extremely fast Safire2™ and Infinite™
F500 microplate readers offer seven different detection modes, saving
valuable time for a wide range of high throughput applications in
the biotech, pharmaceutical, research and cosmetics industries.
These two modular and upgradeable systems provide very reliable
performance with top quality results and are easily integrated into
Tecan’s Freedom EVO® series of liquid handling workstations.
Relax
your reader is a Tecan
www.tecan.com/relax
Cellerity™ is now available
with the MultiChannel Arm™
The Cellerity™, Tecan’s fully automated
cell culture system, is now available with
the MultiChannel Arm™ (MCA) module,
increasing the speed and throughput of
plating cells into microplates for assays.
The MCA module has a 96-channel
pipetting head and can be operated with
washable and disposable tips. The new
option is compatible with lidded and
insert plates, and all standard Cellerity
options, delivering a greater level of
flexibility to your cell culture platform.
5
C L I N I CA L D I AGNOSTICS
Freedom EVOlyzer®
received with great
enthusiasm in Spain
The Freedom EVOlyzer, Tecan’s new
generation of automated ELISA
systems, has proved to be increasingly
popular with laboratories throughout
Spain since being introduced to the
country in July 2006 by Inverness
Medical Iberica (IMI), one of Tecan’s
principle diagnostic distributors
for the area. IMI, formerly part of
Innogenetics Diagnostics, has a long
and proven track record as a distributor
for professional diagnostic products in
Spain, with a strong focus on enzymelinked immunosorbent assays (ELISAs)
for infectious diseases, diagnostics
and autoimmunity. IMI’s customers
range from large hospital laboratories
performing routine testing, to
reference laboratories and testing
laboratories for small- to mediumsized hospitals.
Tecan’s cooperation with IMI started
in the early 1990s and, during the
last decade, IMI has placed a broad
The Freedom EVOlyzer for automated ELISAs
range of Tecan instruments in Spain,
including Genesis RMP™ systems,
Minilysers, Profiblot™ analyzers, Sunrise™
plate readers and Columbus™ plate
washers. IMI is a distributor for nine
international reagent manufacturers and
the company’s combined kit portfolio
consists of more than 300 different tests,
which IMI has successfully automated
using Tecan’s open ELISA platforms such
as the Minilyser and the Genesis RMP.
The addition of the Freedom EVOlyzer has
significantly strengthened IMI’s IVD-D
compliant portfolio.
“We chose to take full advantage of
the large range of customization and
configuration possibilities that the
Freedom EVOlyzer offers, such as being
able to freely combine stainless steel
tips and disposable tips on the same
instrument,” explained Mr Sergio
Gordillo, Automation Product Manager at
IMI. “The instrument’s flexibility for both
hardware and software options means
we are able to satisfy almost all kinds
of ELISA customer with just one single
platform, which has been a key factor
in the Freedom EVOlyzer’s success. We
always deliver these instruments with
a touchscreen monitor, because the
Freedom EVOlyzer software has a
touchscreen compatible interface
available in both Spanish and Catalan,
and this has proven to be a real hit
among our local customers.”
“We have been able to transfer 100% of
our kits to the Freedom EVOlyzer with
excellent results,” continued Mr Gordillo.
“This new system is a more than
worthy successor of Tecan’s Genesis
RMP and its function scope shows a
clear improvement in terms of both
throughput and flexibility.”
“We have relied on Tecan
instrumentation for a good number of
years now and it has proven to be the
right choice. With the Tecan instruments
and our broad range of assays we have
managed to gain a significant market
share of the Spanish market and, with
more than 80 ELISA platforms placed in
Spain, everybody knows that we are a
force to be reckoned with!” claimed Mr
Gordillo. “With the Freedom EVOlyzer we
can now offer superior kits, automated
with superior instruments supported by
superior service. What customer would
ever say no to that?”
CLI N ICAL DIAGNOST I C S
Automated virus
testing in animals
Scientists at the Staatliches Veterinäruntersuchungsamt (National Veterinary Laboratory) Arnsberg
in Germany, responsible for veterinary diagnoses, have chosen Tecan’s Freedom EVO® workstation
for the rapid detection of viruses which cause potentially devastating diseases. The workstation is
already being used to detect the virus responsible for a recent outbreak of bluetongue disease in
cattle, and will soon be used for the detection of the avian influenza virus in local bird populations.
The Staatliches Veterinäruntersuchungsamt is a government
institution in the administrative district
of Arnsberg, North Rhine-Westphalia,
with departments for pathology,
serology, virology and molecular biology
for the diagnosis of animal diseases. It
also investigates food and feed quality,
and performs the detection of pathogens
in foodstuff. Within the organization,
the molecular biology department exists
as a service laboratory to the other
departments by performing molecular
testing for various diseases.
Dr Jochen Kilwinski of the molecular
biology department explained: “Since
September we have been detecting
bluetongue virus RNA in samples from
cattle using Tecan instrumentation,
working in parallel with the serology
department which is detecting
antibodies to bluetongue virus in the
same samples. Bluetongue is an insectborne disease, primarily of sheep, and
there is currently no effective treatment.
It is normally rare in cattle, however, an
outbreak in the Netherlands in August
2006, then spread to the North RhineWestphalia region in autumn. Very
interestingly, the virus detected in this
outbreak belongs to serotype 8, which
normally only occurs in the south of the
Sahara, India, Pakistan, or in central and
South America. Furthermore, serotype
8 is transmitted by biting midge species
of the genus Culicoides, which are not
found in our latitudes.”
“We are using a Freedom EVO 150 to
isolate and purify RNA from blood
samples, to detect the specific RNA
From left to right: Yvonne Kullman, Beatrix Hanstein, Daniela Reckling and Jochen Kilwinski
segment of the bluetongue virus serotype
8 by reverse transcription and real-time
PCR. We obtained the detection protocol
from the Friedrich-Loeffler Institute
in Insel Riems, the National Reference
Laboratory, which is also equipped with
a Tecan instrument. Our Freedom EVO
is working with the Te-VacS™ vacuum
manifold, with a 96-well format to handle
a large throughput of samples.”
Dr Kilwinski is delighted with the
results he has been getting from the
Freedom EVO. “During initial testing,
there was 100% agreement between
the preparations made by the Freedom
EVO and our manual preparations.
Furthermore, the automated preparation
was shown to be in good accordance
with the handmade preparations while
participating in a national bluetongue
virus ring trial. We have a lot of
experience with manual handling, so this
gave us complete confidence in Tecan’s
automated system. The performance
of the Freedom EVO is excellent with
no reliability problems, and we would
not have been able to handle our
current throughput manually. We are
very happy to have this instrument.
I strongly feel that other institutions
should also have the opportunity to
carry out their investigations with a
workstation like this.”
With the flexibility of the Freedom
EVO, Dr Kilwinski’s workstation is not
restricted to bluetongue virus detection.
“We are also responsible for the
detection of avian influenza in wild bird
populations within the administrative
district of Arnsberg. The protocol for
the detection of the avian influenza
virus has already been implemented
on the Freedom EVO, and it is very easy
to switch from testing for bluetongue
virus to avian flu. We are now waiting
for spring, when the migration of birds
starts again, and we are ready to use
the workstation to test for avian flu
in local bird populations, should the
need arise.”
Q ualit y A ss u rance
Working for
ISO accreditation
around the world
Following the successful ISO certification
of our manufacturing sites, work
is already under way to implement
similar high standards in all of Tecan’s
organizations around the world. The first
sales organization, Tecan France, has now
been certified according to ISO 13485,
an international standard intended for
medical device companies.
Implementation of this quality
management system means that all sales
and service activities throughout our
sales organizations will be structured in
a similar way, and customers everywhere
will receive the same consistently high
standard of service and support from
Tecan. As with our manufacturing
facilities, we have chosen to work with
TÜV Product Service, a well-known global
notified body with a particular expertise
in the medical device business and a
strong reputation for high standards.
Günter Weisshaar, Tecan’s Executive
Vice President Global Quality and
Regulatory Compliance, is leading the
implementation at a global level, and
explained: “Implementing the right
structure and procedures into our
market units will help us to become
more customer oriented and to respond
in a more timely and efficient manner.
An important benefit will be a change
in the way we handle information from
our customers; whether in the form of
a complaint or a product suggestion, we
will be able to handle this information in
a far better way.”
Erika Hildebrandt, Quality Manager for
Tecan Europe, led the project in France
in close association with Denis Coulet,
Business Manager for Tecan France. She
explained: “Implementing ISO 13485
helped us to document successful
processes already in place, and to improve
other procedures. The main focus of all
the changes is to serve our customers
in a better and more professional way
and, only a few months on from the
implementation in France, we can already
see the advantages. The everyday running
of Tecan France has become more precise
and our customers are already feeling
the benefits. Installations, maintenance
and overall customer service have all
improved as we are able to record and
track customers’ requests more easily
and respond more quickly. For the long
term, too, we have refined our methods
of collecting customer information and
feeding this back to our manufacturing
and R&D sites so that it can be fed into
new product designs where appropriate.”
The quality system now established in
France will be systematically implemented
into all Tecan’s sales organizations and we
look forward to a time very soon when
our customers around the world will be
able to enjoy the same advantages as our
customers in France.
A P P L I C AT I O N B I O P H A R M A
High throughput nucleofection®
with the Freedom EVO® workstation
and amaxa 96-well Shuttle®
Tecan and amaxa GmbH have successfully integrated the amaxa Nucleofector® 96-well Shuttle®
System with the Tecan Freedom EVO® liquid handling workstation, allowing fully automated,
high throughput transfection of primary cells and difficult-to-transfect cell lines, including
non-dividing neurons and T cells, with siRNA, DNA and other substrates. The collaboration was
originally instigated for Dr Claudia Merz at Bayer Schering Pharma AG, Germany, who needed fully
automated, high throughput transfection of immune system cells for RNAi-based
screening experiments.
Scientists routinely use transfection
techniques to insert DNA or RNA into
cells and modify the expression levels of
a certain gene or genes. This approach
is often used for functional studies of
genes, such as RNA interference (RNAi)
to silence gene expression; for cell-based
manufacturing of therapeutic products
(eg. antibodies, hormones or vaccines);
and for structural studies of proteins and
amplification of DNA or RNA in cells for
purification. Many of these applications
require high throughput transfection in
combination with primary cells or nondividing cell lines that are notoriously
difficult to transfect with standard
lipofection approaches.
The Nucleofector® technology from
amaxa, a German biotechnology
company, has been specifically designed
for difficult-to-transfect cell lines and
primary cells, including stem cells,
T lymphocytes and non-dividing neurons.
The technology uses electroporation
instead of lipofection, where electrical
currents cause the gene of interest to be
transferred directly into the cell and even
the cell’s nucleus. This quick and simple
approach is the only non-viral method to
achieve very high transfection efficiencies
while maintaining high survival rates in
such cells, with up to 80% efficiencies in
human T cells, for example, compared
with 1-3% or fewer usually transfected by
lipofection. As nucleofection causes the
DNA to directly enter the nucleus, this
10
The entire nucleofection process is
controlled by the Freedom EVOware®
software, using an interface provided
by amaxa. Nucleofection conditions
for the 96-well Shuttle® are defined in
the amaxa software and the resulting
parameter files are uploaded from
Freedom EVOware and executed on the
96-well Shuttle®.
method does not require cell division, and
expression of the transfected gene can be
analyzed shortly after nucleofection. For
some cell types this can be within as little
as two hours, rather than the standard 24
to 48 hours.
Integrated cooling and heating elements.
Transportation of amaxa’s Nucleocuvette™
plate to the 96well Shuttle.
Tecan and amaxa have teamed up to
integrate amaxa’s Nucleofector® 96-well
Shuttle® System with Tecan’s Freedom
EVO® liquid handling workstation. All
necessary steps for transfection can be
automated, including cell harvesting,
diluting and plating; DNA/RNA
normalization; preparation of reagent
mixes; resuspension of cells and
substrates in Nucleofector® solution; the
nucleofection process; and analysis of
transfection results. Nucleofection occurs
in disposable, 96-well Nucleocuvette™
modules, which consist of an innovative
conductive polymer electrode material
that ensures no metal ions are released
into the cell suspension during
transfection. The system uses identical
transfection parameters for any nucleic
acid substrate, so that DNA vectors, such
as expression plasmids, and RNAi vectors,
including short hairpin (sh)RNA vectors or
short interfering (si)RNA duplexes, can be
transfected using the same protocol.
“Many easily-transfected cell lines are
quite different from primary cells of the
human body so are not applicable for
most research and drug development
studies,” explained Andreas Schroers,
Product Manager at amaxa. “The
integration enables, for the first time,
transfection of medically relevant primary
cells in a fully automated environment,
providing a valuable tool for research and
drug discovery studies.”
The collaboration originally began in
response to a request from Dr Claudia
Merz, scientist in Target Discovery at
Bayer Schering Pharma AG in Berlin,
Germany. Dr Merz had been performing
lipofection using a Freedom EVO
workstation for some time, and needed
to expand the set-up for large scale,
automated nucleofection of human
T lymphocytes and the Jurkat lymphocyte
cell line using focused siRNA libraries.
These small libraries have between
200 and 800 siRNAs that are applied
in triplicate, requiring multiple 96well Nucleocuvette™ plates to be
completed each day. Dr Merz’s integrated
system automates the complete plate
preparation, nucleofection and replating
of nucleofected cells for one 96-well plate
in about 40 minutes, and twelve 96well plates can be processed in one day,
equating to 1,152 samples. A Freedom EVO
200 workstation is used for all additional
steps of the RNAi studies, such as mRNA
knockdown analysis and analysis of
phenotypic changes of the knocked down
cells.
Come and see the amaxa integration on
show on Tecan’s booth 311 at the Society
for Biomolecular Screening exhibition in
Montreal, Canada, April 2007.
96-well Nucleofection
Lipid-based reagent L
100 % transfection efficiency
90
80
70
60
50
40
30
20
10
0
HL-60
Jurkat
K562
U937
Nucleofection versus lipofection of suspension cells
Cells were transfected with (pmaxGFP™). The efficiency was
measured on a BD FACSCalibur™ 24h post transfection.
% transfection efficiency
“We are very excited and pleased that
the final installation of the integrated
system into our laboratory was achieved
so smoothly, including setting up the
software protocols to run the 96-well
transfections,” said Dr Merz. “Our
protocols for manual transfection of
primary human T cells or Jurkat cells, for
example, have been fully adapted for high
throughput nucleofection on the Freedom
EVO workstation. We can now run RNAi
screens using difficult-to-transfect cells,
primary cells or suspension cells in an
automated fashion, which will enhance
and facilitate our target discovery efforts
in dermatology or immunology, for
example.”
11
100
90
80
70
60
50
40
30
20
10
0
Column 1
Column 2
Column 3
Column 4
Reproducible intra-plate transfection efficiency in nucleofected
Jurkat E6-1 cells (ATCC® TIB-152™)
Analysis was performed on a BD FACSCalibur™ 24h post
Nucleofection. The transfection efficiency of each well is shown
per column of a 96-well Nucleocuvette Module. Column 4 contained
two control samples (no pulse, no plasmid). (Data kindly provided
by C. Merz. Bayer Schering Pharma AG, Berlin)
100 relative expression (% pulse only, sample C)
50
Not for clinical use or blood and plasma screening.
The Nucleofector 96-well Shuttle System
and the 96-well Nucleocuvette plates and
modules are covered by patent and/or patentpending rights owned by amaxa. PmaxGFP is a
trademark of amaxa.
BD FACSCalibur is a trademark of BD Biosciences.
ATCC is a registered trademark and TIB-152 is a
trademark of American Type Culture Collection.
40
30
20
10
0
1
3
5
7
9
11
13
15
C
siRNA-mediated depletion of vimentin in human T-cells
Knockdown on mRNA level measured by qRT-PCR. 15 samples compared to control (C)
set to 100% (Data kindly provided by C. Merz, Bayer Schering Pharma AG, Berlin)
12
L IQ U I D H A N DLI NG AN D ROB OTICS
The target of personalized
medicine moves ever closer
Researchers at Genomic Health Inc., California, are using several Freedom EVO® 200 workstations
and a Genesis platform to automate the development and routine screening of gene panels for
predicting the risk of recurrence and response to treatment of breast cancer.
Scientists now firmly believe that
an individual’s genetic makeup has
a strong influence on susceptibility
to disease onset or recurrence and
response to different treatments, and,
as a consequence, understanding these
genetic variations is seen as central to
creating personalized drugs, tailored to
the individual, with greater efficacy, safety
and fewer adverse side-effects.
Screening genomes for these variations
is logistically awesome as there are
potentially many candidate genes
that are likely to be involved. The focus
of research and development at life
science and clinical diagnostics company
Genomic Health Inc. is to do just that
and current efforts are concentrated on
developing a gene panel which serves as
an initial screen of many genes.
In 2004, Genomic Health launched
the Oncotype DX™, the first diagnostic
multi-gene expression test commercially
available that has clinical evidence
validating its ability to quantify the
likelihood of breast cancer recurrence
in women with newly diagnosed, early
stage, node-negative, estrogen receptorpositive breast cancer. The Oncotype DX™
assay analyzes the expression of a panel
of 21 genes from a tumor specimen using
RT-PCR, and the results are presented as a
Recurrence Score™ that estimates the risk
of cancer recurrence within ten years, on
a scale from 0 to 100. These scores
are more accurate and reproducible than
the variables that clinicians currently
rely on, like tumor size and grade, and
patients’ ages.
The assay has five steps, starting with
manual RNA extraction from formalinfixed, paraffin-embedded tumor tissue
(FPET) samples. All the subsequent
steps are automated, from RNA
quantification by fluorescence assay,
to a quantitative PCR assay that checks
for DNA contamination, and reverse
transcription (RT) which produces
complementary DNA from the mRNA.
The final step is quantitative TaqMan®
RT-PCR using the complementary DNA as
LIQU I D HAN DLI NG AN D ROB OT I C S
the template in 384-well plates, with the
expression of each of 16 genes measured
in triplicate and then normalized
relative to a set of five reference genes.
RT-PCR is highly reproducible as it is
sensitive, specific, unaffected by inherent
variability in the original tumor blocks,
and amenable to high throughput and
real-time quantification, with successful
amplification even from degraded
starting RNA.
The work to develop the Oncotype DX™
assay down to a much more manageable
21 genes has relied on three independent
clinical studies and the assay has been
validated in a large, independent multicenter clinical trial (NSABP Study B-14)
and in a large population-based
case-control study in breast cancer
patients at the Northern California Kaiser
Permanente. The magnitude
of these and potentially more studies in
the future made the use of automation
in the development work essential, as
Jay Snable, Director of Process
Automation, explained:
“We needed automated systems for
our ongoing assay development and,
although we chose Tecan for several
reasons, the primary reason was the
highly flexible technology, offering so
many accessories that we can basically
do whatever we need to. As a clinical
reference laboratory, we receive the
patient samples in tubes, and Tecan
provided the best solutions for handling
all the different formats we need,
from single tubes through to 384-well
microplates, in a highly integrated
system with quality controls all the way.
It incorporates integral barcode scanning
and identification, enabling tracking of
individual samples through all the
assay steps.”
“A big labor-saving feature of the platform
comes from the barcode information that
the system generates during assembly
of samples into assay plates. We can
put samples on in any order; they are
scanned and information is sent to our
LIMS where the tube and plate barcodes
are married, eliminating manual batching
steps. This was just one of the benefits
coming from the successful integration
of the equipment with our LIMS. For
example, we also use the LIMS for
normalization. Tubes are placed on the
robot and scanned. The resulting file is
exported to the LIMS and a Gemini™
script for adjusting sample volumes and
concentrations is generated and sent
back to the platform. The samples are
then normalized for use in some of the
intermediate steps of the Oncotype
DX™ assay, the genomic DNA assay and
reverse transcription.”
Most of the Tecan workstations at
Genomic Health are configured with
Tecan’s Multichannel Pipettor, Te-MO™.
The PCRs are set up on the Freedom
EVO workstations using Te-MO 3/5
multipipettors with 96-channel pipetting
heads (Te-MO 96). These workstations
are also equipped with a PosID™ System
for automated barcode reading. Applied
Biosystems’ 384-well thermocyclers
(7900HT Fast Real-Time PCR systems)
are used for the RT-PCR and resulting
quantification. The data are reduced
and analyzed using custom software
developed in-house and, after passing
a series of quality control steps, a
Recurrence Score™ is calculated.
A report is generated, reviewed by clinical
staff, and released to doctors who advise
the patients.
Jay concluded: “In 2005, we ran about
7,000 RNA samples in the clinical
laboratory with a targeted turnaround
time of 10 to 14 days per sample, including
review and quality control processes. We
continued to scale up through 2006 and
processed approximately 13,500 patient
samples. Genomic Health is also now
scaling up the development process. For
example, we are now looking at 761 genes
in an initial screen to create a gene profile
for an assay to test for risk of recurrence
of another cancer type, and this is where
the Tecan technology will prove especially
useful for the high throughput we need
to achieve.”
From left to right: Jay Snable, Darrin Chan,
Keith Volk, Kenneth Hoyt, Anhthu Nguyen
and Ian Storer from Genomic Health.
Not all options discussed in this article have been cleared for use in all markets. Contact your local
sales office for specific information.
Oncotype DX and Recurrence Score are trademarks of Genomic Health, Inc.
TaqMan is a registered trademark of Roche Molecular Systems, Inc.
13
14
FO R E N S I C S
Genetic fingerprinting
of crime scene samples
Forensic scientists at the Landeskriminalamt police office in Stuttgart, Germany, have
commissioned a semi-automated sample processing system from Tecan to handle about
20,000 crime scene samples it receives annually for genetic fingerprinting. The specially
designed system includes one MiniPrep™ and two Tecan Freedom EVO® liquid handling
workstations. In addition, the system includes an integrated thermal cycler, real-time
PCR and capillary electrophoresis systems from Applied Biosystems.
The Landeskriminalamt (LKA) is the
central criminal police office for the
federal state of Baden-Württemberg
(BW), in South-West Germany, and its
forensic services division is responsible
for investigating most of the biological
crime scene samples received from the
deferring police departments in the
state. Approximately 20,000 samples
per year come from crimes of all types,
including murders, thefts, sex crimes
and vandalism. Forensic scientists need
to be able to process the complete range
of sample materials. For each sample,
genomic DNA is extracted from the
cells, purified and quantified prior to
amplification of short tandem repeats
(STRs) by PCR. Sequencers are then used
for fragment analysis of STRs, using
capillary electrophoresis, and a genetic
fingerprint is formed.
In 2005, Dr Kai Vollack and his colleagues
Dr Barbara Haak and Dr Ralph Schwenzer,
the automation integration team at the
LKA-BW, started looking for a semiautomated method for processing
their samples in order to cope with the
demanding workload. The laboratory had
a number of stringent requirements that
the automated system needed to meet. It
needed to be able to handle a wide range
of biological sample types and volumes;
to provide the maximum DNA yield and
highest analysis sensitivities; to ensure
minimal risk of DNA contamination;
to incorporate barcode-based sample
identification; and to integrate real-time
PCR using human-specific primers and
performing PCR of STR DNA sequences.
The members of the automation
integration team of the LKA-BW
(from left to right: Dr Barbara
Haak, Dr Werner Pflug, Anja
Gruber, Angelika Jelinski, Dr
Ralph Schwenzer, Jessica Loidl,
Marga Kugler and Dr Kai Vollack).
Another requirement of the LKA was
based on the pioneering work, started
in the early nineties by Dr Werner Pflug,
the head of the DNA division, using
evidence from perpetrators’ skin cells for
DNA profiling. “When a person touches
an object, he leaves single particles from
his skin on that object,” explained Dr
Vollack. “It was originally thought that
these particles were dead cells with
totally degraded DNA but we now know
that this is not the case and we often
have enough DNA in these particles to
create a complete genetic fingerprint.”
For this reason, it was particularly
important that the set-up could
separate epithelial cell samples that do
not contain any DNA from those that
do contain DNA at an early stage within
the processing workflow. No available,
off-the-shelf system could meet these
requirements so the project went to
public tender and Tecan was chosen for
a number of reasons.
“The main advantage of choosing
Tecan was the flexibility of the whole
system,” explained Dr Vollack. “We
needed a platform that was open to
new developments and compatible with
devices and reagent kits from different
companies. We had seen a Tecan system
in action at Professor Scheithauer’s
laboratory at the department of legal
medicine, Medizinische Universität
Innsbruck, and we appreciated its
outstanding performance, particularly
with the sample identification provided
by the integrated barcode reader and the
automated data exchange with the LIMS.
We were also invited by Tecan to see the
automated genetic sample processing
system that was recently created for the
South African Police Service and we were
very impressed with this.”
The forensic science division at the LKABW now has its own fully operational
system provided by Tecan which has
semi-automated sample analysis. The
first step of the process involves manual
separation of the cell lysate from the stain
carrier, which could be a skin particle,
piece of tissue or swab, for example. The
lysate is transferred to a Freedom EVO
150 workstation, which is equipped with
FOR E N S I C S
Manual separation of the cell lysates
from the stain carriers and preparation
of the crime scene samples for the
ChargeSwitch® Technology magnetic
bead extraction of genomic DNA.
an 8-channel liquid handling (LiHa) arm,
two Te-MagS™ magnetic bead separation
modules and a PosID™ barcode sample
identification device. Samples undergo
magnetic bead-based DNA extraction
and purification using the ChargeSwitch®
Technology from Invitrogen and eluted
DNA is pipetted into 96-well PCR plates.
These plates are transferred to the second
Freedom EVO 150 workstation for the PCR
set-up for DNA quantification using the
Quantifiler® Human DNA Quantification
Kits from Applied Biosystems. Samples
undergo real-time PCR using an
integrated Applied Biosystems 7500 RealTime PCR System and data are exported
to the LIMS and imported into the second
Freedom EVO workstation for STR PCR
set-up of samples containing sufficient
DNA, then sent to an Applied Biosystems
9600 Thermal Cycler. The samples are
prepared for capillary electrophoresis by
the third Tecan liquid handling system,
a single-channel MiniPrep 75, and finally
subjected to capillary electrophoresis
with a 16-capillary Applied Biosystems
3130xl Genetic Analyzer.
The system includes a number of extra
safety precautions in the workflow,
including several that are thought to
avoid cross-contamination of samples.
For example, the liquid handling arms
do not travel across open wells or tubes
other than their targets, and using the
Te-MagS module instead of a shaking
device for mixing prevents the formation
of potentially contaminating aerosols.
The forensics division at LKA-BW has a
‘four-eye’ policy, where a second member
of staff must confirm every sample
transfer, which is labor-intensive as well
as time-consuming, but incorporating
Tecan’s automated sample identification
processes within the workflow has helped
to free up personnel for other tasks.
“We are really happy with the system,
it is very sensitive and we have already
tested a wide range of possible samples,
including the more difficult skin particle
samples,” said Dr Vollack. “The automated
processing also reduces operational error
and, therefore, improves the quality of
our data.”
Preparation of single epithelial particles
from the butt of a pistol used as a
murder weapon.
ChargeSwitch is a registered trademark of
Invitrogen.
Quantifiler is a registered trademark of Applera
Corporation or its subsidiaries in the US and/or
certain other countries.
The Tecan Freedom EVO 150
liquid handling workstation
with two Te-MagS magnetic
bead separation modules for
DNA extraction of crime scene
samples using the ChargeSwitch®
Technology from Invitrogen.
15
16
M I C ROA R R AyS
Profiling the expression patterns
of hundreds of microRNAs with the
HS 800™ Pro Hybridization Station
Exiqon is a Danish biotechnology company that provides products and services for the life sciences,
research and drug discovery industries all over the world. It specializes in microRNA (miRNA) analysis
and produces unique, locked nucleic acid (LNA™)-based high affinity arrays for miRNA profiling. Tecan
HS 800™ Pro Hybridization Stations were used during the development of these arrays and Exiqon
now relies on the HS 800 Pro systems to provide miRNA profiling and analysis services.
MicroRNAs have recently generated
a great deal of interest and are
now recognized as important posttranscriptional regulators of gene
expression in a variety of biological
functions (reviewed in 1-). However,
the actual functions of miRNAs and the
mechanisms by which they are controlled
remain unclear. Many organisms
express hundreds of miRNAs that vary in
abundance across cell and tissue types.
The total number of miRNAs in vertebrate
genomes is currently unknown, but
hundreds have already been isolated
and the final numbers may reach over
1,000. A great deal of research remains
to be carried out into these molecules,
including their possible roles in human
diseases and potential for novel
therapeutic tools.
Exiqon, a biotechnology company based
in Copenhagen, Denmark and Woburn,
USA, produces the miRCURY™ LNA
microarrays, which were specifically
developed for miRNA expression
profiling. Using a high affinity nucleotide
analogue, locked nucleic acid (LNA™),
Exiqon creates oligonucleotide probes
with greatly increased affinity for their
complementary targets, due to the
increased melting temperature (Tm). This
has proved to be particularly useful for
detecting short nucleic acid targets such
as miRNAs, as standard oligonucleotide
technologies have insufficient specificity
and affinity for these molecules. The
miRCURY™ LNA arrays have almost 1,500
LNA™-based capture probes, making
it possible for researchers to profile all
miRNAs from vertebrates, invertebrates,
M I C R OA R R AyS
plants and viruses (as registered in the
miRBase 8.1 database at The Wellcome
Trust Sanger Institute, UK) and to crossprofile between different species. The
microarrays also contain a number of
capture probes (miRPlus™ probes) for
detection of miRNAs that are not included
in miRBase. The microarrays have low
sample usage requirements, being able to
work with as little as 1 µg total RNA, and
do not require miRNA enrichment.
Exiqon’s miRCURY™ LNA arrays were
developed using a Tecan HS 800 Pro
Hybridization Station, which offers fully
automated, highly sensitive microarray
processing with high reproducibility
and flexibility. “We started using the HS
800 Pro during the development of
our array products because we needed
an automated system for hybridizing
and washing the slides,” explained
Ina Dahlsveen, Product Manager for
Services at Exiqon. “It is very important
to have as little variability between
slides as possible, and using the HS 800
Pro improves our reproducibility and
minimizes technical issues that can arise
during manual hybridization. Although
our slides are compatible with the
manual systems available, these are less
efficient for processing large numbers of
slides.”
In May 2006, Exiqon started providing
miRCURY™ LNA array profiling services, as
Peter Roberts, Brand Manager at Exiqon,
explained: “The services were actually
set up in response to specific demand.
After we launched our microarrays,
we were getting at least one or two
enquiries every week asking if we could
run an analysis service, despite the fact
that we never advertised one. So when
we set the services up, it was a natural
progression for us to incorporate the
Tecan hybridization stations with them.”
samples from plants, zebrafish, C. elegans,
domestic cow and so on – there are
customers out there working on all kinds
of wonderful organisms. Our turnaround
time is variable depending on the size of
the project; at the moment, an average
project involves about 10 to 12 slides
and takes about two weeks from when
we receive the samples until we deliver
the results. Our services include data
analysis, depending on the customer’s
requirements. Every project is different
so we try to be as flexible as possible and
customize our services for each customer
and their samples.”
“The arrays work very well with the Tecan
hybridization stations, there is very good
compatibility between them,” added
Peter. “We have had a few instances
where customers that are using our
arrays for the first time come across
some technical problems but, by taking
a Tecan hybridization station on-site and
running the arrays with the customer, the
problems have been solved.”
The compatibility is so convincing
that Exiqon’s US office in Woburn,
Massachusetts, routinely sends Tecan
hybridization stations to its new
miRCURY™ LNA customers to aid them in
setting up new array procedures. Mikkel
Nørholm, Manager of Technical Sales
Support at Exiqon in the US, explained:
“When a laboratory starts using a new
technology, it normally takes some time
to get everything working perfectly.
During that process it’s really helpful
to have a gold standard, something
that you know works. Combining a
Tecan hybridization station with our
predeveloped miRCURY™ LNA protocol
means that even for inexperienced users,
the procedures work straight away.”
The program for Exiqon’s automated
microarray hybridization protocol was
originally developed in collaboration
with Erasmus MC, based in Holland.
“The reproducibility you can get with
the hybridization stations, even across
different laboratories, is so high that
we could simply install the program
exchanged by e-mail and it worked
instantly!” said Mikkel. “This is also a
major benefit for our customers as they
can start on their experiments much
more quickly after receiving the program
from Exiqon electronically.”
References:
1. Bartel DP (200). MicroRNAs: genomics,
biogenesis, mechanism, and function.
Cell 116(2): 281-297
2. Shivdasani RA (2006). MicroRNAs:
regulators of gene expression and cell
differentiation. Blood 108: 66-65
3. Wang Y et al. (2007). MicroRNA: past
and present. Frontiers in Bioscience 12:
216-229
LNA, miRCURY and miRPlus are trademarks of
Exiqon A/S, Vedbaek, Denmark.
“Obviously with services you also
need high throughput, as well as good
reproducibility,” Ina continued. “We soon
acquired a second HS 800 Pro system,
and we will get a third station in the near
future. Most of the samples we receive
are human, but we also get many samples
from mouse and rat, and our newest array
contains capture probes for miRNA from
all types of organisms. We have received
17
18
LSIQ
D HRAEN DLI NG AN D ROB OTICS
O FUTIWA
Engineering new
biomaterials
for human implants
Scientists at the University of Applied
Sciences in Zurich, Switzerland, are
collaborating with Tecan to optimize
automated liquid handling systems for
cell and tissue culture. The Cell Biology
Division at the University’s Institute
of Chemistry and Biotechnology (ICB)
specializes in developing cell culture
technologies and tissue engineering,
including the in vitro development and
investigation of biomaterials for bone,
cartilage, intervertebral discs, teeth
and other connective tissue implants.
It is essential that these materials are
tested to high standards of quality with
minimal variation, so the group has been
developing automated tissue culture
methods using a Tecan Freedom EVO®
liquid handling workstation as part of
their research activities.
Professor Dr Ursula Graf-Hausner, Group
Leader of the Cell Biology Division, said:
“We are collaborating with industrial
partners who develop biomaterials, such
as titanium-based products, as potential
foreign implants. We begin by carrying
out in vitro studies of these biomaterials
and, if the results of our preliminary
studies are successful, then the materials
proceed to animal model studies and,
eventually, to clinical applications. Some
of the technologies we use are still in the
early stages of development but I believe
an essential factor for the success of
any method is that it can be automated.
Reproducibility is an extremely important
part of reaching our clinical goals when
working with these materials, and we
have started a very good collaboration
with Tecan to develop these methods.”
The Cell Biology group has been
evaluating the capabilities of the
Freedom EVO workstation to perform
fully automated cell culture, including
cell harvesting and plating, for several
different cell lines. The Freedom EVO is
equipped with a FlaskFlipper™ module
for handling the automation-friendly
Corning® RoboFlask™ vessels and an
Infinite™ F200 luminescence reader for
analyzing the cells. It is also configured
with a modified, 8-channel liquid
handling arm equipped with 5 ml syringes
that is capable of both rapid bulk media
dispensing and precise low-volume
pipetting for reagents, as well as a robotic
manipulator arm, and a Te-Shake™ robotic
shaker and stirrer.
The group uses a variety of mammalian
cell lines and human primary cells for its
research and has been testing the
Freedom EVO’s ability to handle cell types
with different adhesion and aggregation
characteristics, including mouse
fibroblasts (moderate adherence and
aggregation), human osteogenic sarcoma
(SaOS-2) cells (strong culture flask
attachment and aggregation) and human
breast adenocarcinoma (MCF-7) cells
(weak attachment to flasks, moderate
aggregation). The harvesting efficiencies
of the workstation and the viabilities of
the cultured cells were compared to
manually obtained results, and it was
found that the automated methods
achieved comparable or better
reproducibility and viability of cells than
manual methods1 (see Figure 1).
Dr Stephanie Mathes, research scientist in
the Cell Biology group, has been running
the Freedom EVO workstation, and said:
“The flexibility of the Freedom EVO is
really positive and specific design features
can easily be added to suit a customer’s
application. I particularly like the Freedom
EVOware® software; it is very easy to
operate and you can simply drag and drop
icons to create a whole new automated
procedure. We have plans for more
projects now that we’ve seen the true
flexibility of Tecan’s workstation and all
its possibilities.”
“The collaboration we have with Tecan is
very close and we have really been able to
share our knowledge and experiences,”
Professor Graf-Hausner added. “For
example, in some instances we have
achieved better data by using slightly
different volumes of reagents and have
passed this on to Tecan for optimizing the
system.” The team has recently developed
a range of complete, automated cell
harvesting and plating protocols that
will accommodate different operating
conditions, such as the time required for
the cells to detach from culture flasks,
100
8
7
99
6
Cell viability (%)
“The automated system saves us a lot of
time in preparing cells, not least because
we can leave it to run unattended
over the weekends and during holiday
periods,” Professor Graf-Hausner
continued. “This is a huge advantage
for members of our team, including
several students, who would normally be
manually pipetting the cells, as they can
now spend their time doing much more
interesting and sophisticated tasks. The
collaboration as a whole has given them
an excellent insight into the early steps
of process development, and the system
itself shows what cell culture technology
and tissue engineering research might
involve in the future and how new
technology will really make our lives
much easier in the laboratory.”
From left to right: Lorenz Wohlwend, Stephanie Mathes and Ursula Graf-Hausner.
Cell number/RoboFlask (mio)
to suit a variety of cell types. The results
generated are easily transferable to other
Tecan cell culture equipment such as the
Cellerity™.
5
4
3
2
Cell number
Robot
Reference:
1. Graf-Hausner U, Mathes S, Baumgartner
S, Mueller U (2007). Automated
processing of cell cultures on the Tecan
Freedom EVO robotic workstation. Poster
presented at ALA 2007.
Further details of the study are available
at www.tecan.com
Corning is a registered trademark and RoboFlask
is a trademark of Corning, Inc.
Not all applications discussed in this article have
been cleared for use in all markets. Contact your
local sales office for specific information.
97
96
1
0
98
95
Cell viability
Manual
Figure 1
SaOS-2 cells were harvested manually and
automatically by a Freedom EVO workstation equipped
with a FlaskFlipper and Corning® RoboFlask™ vessels.
Cell number and viability were compared. Repeatability
of the process and viability of the harvested cells were
higher for the automated process.
19
20
A P P L I CAT I O N B IOPHARM A
Plasma technology for fast,
effective tip cleaning
Tecan has teamed up with Cerionx Inc., the sole provider of automated pipette tip cleaning systems
using ‘cold’ plasma, in an exclusive global distribution agreement for the supply and marketing of
the Cerionx 8-channel TipCharger™ System.
It is estimated that over 99% of all matter
in the known universe is in the form of
plasma – lightning and flames are the
only two naturally occurring forms on
earth. However, through advancements
in physics and chemistry as well as
the harnessing of alternating currents,
the breadth of man-made plasmas
continues to widen. Pennsylvania-based
company Cerionx uses a man-made,
atmospheric pressure ‘cold’ plasma in its
unique TipCharger™ System to rapidly
and thoroughly clean pipette tips more
efficiently than traditional solvent-based
methods.
Atmospheric plasma begins with the
capturing of free electrons to charged
dielectric plates. Through changes in
polarity, these electrons are forced to
mobilize between a set of plates resulting
Figure 1: Generation of atmospheric plasma
and the subsequent ionization/activation of
room air.
in an electron avalanche (Figure 1).
By drawing air in between these plates
and through the electron field, the
components of air are bombarded with
electrons, causing the fracturing and/or
activation of CO2, H2O, O2, and N2. The
result is a ‘cold’ or ‘non-thermal’ plasma
field where most of the input energy is
channeled to the electron component of
the plasma while the resulting ions and
neutral components remain at or near
room temperature. In addition, highly
energetic metastable atomic species
are generated. These species have the
appropriate number of electrons, but are
displaced to higher shells and exhibit high
reactivity and some selectivity in their
reaction capabilities. The plasma ions and
metastable atomic species react rapidly
with biologicals and organic solvents to
facilitate their breakdown and eventual
removal (Figures 2 and 3).
The patented cold plasma technology
developed by Cerionx is quite unique and,
although plasma has been used in other
cleaning applications, the TipCharger™
is the first to apply it to pipette tip
cleaning in the laboratory environment.
In contrast to the conventional solventbased cleaning methods by dilution,
TipCharger™ generates no solid or liquid
waste, eliminates the use of bleach
washes and generally speeds up current
wash protocols.
The collaboration between Tecan
and Cerionx began in the summer of
2005, when Mr Paul Hensley, founder
of Cerionx, visited the Tecan facilities
with an early model of the TipCharger™
system. Tecan’s R&D laboratory tested
the unit thoroughly to fully assess its
potential with Freedom EVO® liquid
handling workstations, and an exclusive
global distribution agreement for
Tecan to market the Cerionx 8-channel
TipCharger™ System was finalized in
December 2006.
Ms Tia Smallwood, Director of Marketing
at Cerionx, elaborated: “We knew
that TipCharger™ would provide a
manufacturer of liquid handling
workstations with significant added value
and help differentiation in a crowded
market. We approached Tecan as the
worldwide leader recognizing they could
position TipCharger™ to maximum
advantage. We really believe that the sum
of these two products is greater than
each of the parts.”
APPLICATION
B IOPHA RNMTS
A
COMPONA
Figure 3: Artist’s impression of plasma cleaning action of pipette tips in the TipCharger™ cleaning station.
Even before the agreement was signed,
Tecan and Cerionx worked closely
together to provide several customers
with the first systems to combine the
TipCharger™ with Tecan instruments.
One of the early users was Transnetyx
Inc., a genotyping service provider in
Memphis, Tennessee. Mr Bryan Rushing,
Automation Engineer at Transnetyx,
explained: “We had been using a
bleach solution rinse protocol on our six
Tecan Freedom EVO and Genesis RWS™
workstations but, by switching to the
TipCharger™, we found that our protocol
achieved a 40% reduction in processing
time. Processing speed is critical to
our business. We guarantee delivery of
our results in 24 or 72 hours, achieving
99.973% accuracy.”
In the future, Cerionx is planning to
validate the TipCharger™ for clinical
environments, making the technology
available to even more Tecan users, and
new formats of the technology are also
under development. Mr Robillard said:
“Tecan is a great partner for us; we work
in very similar ways. Both companies are
aligned around the notion of proving
technologies one application at a time,
and are committed to development
through proof of application. Tecan is a
tremendous partner to work with and,
at Cerionx, we are really excited to be
working together. We hope and expect
the collaboration to lead to a very active
exchange in the future.”
TipCharger is a trademark of Cerionx Inc.
Mr Mark Robillard, Vice President of Sales
and Marketing of Cerionx, added: “This
early proof of concept demonstrated the
good technical fit of the TipCharger™
with Tecan workstations, and the
experience confirmed our belief that the
genotyping, genomics and like markets
offer extensive opportunity.”
Figure 2: Plasma cleaning action in and
around a pipette tip.
21
22
L IQ U I D H A N DLI NG AN D ROB OTICS
Building a cattle
DNA database
in Japan
Over the last decade, the Livestock Improvement Association of Japan (LIAJ) has developed
a DNA and beef sample database capable of parentage discrimination analysis for pedigree
registration and individual identification of every head of cattle in Japan. To handle the
large number of samples involved, scientists at LIAJ have chosen Tecan’s Freedom EVO® 200
workstation, with barcoded sample identification, for the automated extraction of DNA.
Since its establishment in 1965, LIAJ
has developed its activities, with the
improvement of cattle as the primary
objective. Today, the association continues
to work on livestock improvement, using
various methodologies to support the
production of superior breeding stock
and to develop efficient methods for the
individual identification of animals. The
Maebashi Institute of Animal Science,
LIAJ’s livestock improvement technology
laboratory in Gunma, Japan, was opened
in 1977 to carry out blood typing and
artificial fertilization research in cattle.
The institute has since expanded to
handle embryo transplantation, sperm
sorting, as well as all DNA-based testing
such as parentage testing, testing for
hereditary diseases and individual
identification. In addition to the various
inspection businesses, the Maebashi
Institute is involved in research work,
including quantitative trait loci (QTL)
analysis of Japanese varieties of cattle
using DNA markers, in collaboration with
other research institutions.
As a technical scientist in the Division
of Animal Genetics at the Maebashi
Institute, Dr Kazuhito Kurogi is in charge
of the individual identification and
parentage discrimination analysis. Dr
Kurogi explained: “In Japan, a blood
typing-based cattle identification
database was created in the 1970s, which
LIAJ took over from the Japan Holstein
Registration Association in 1978. The work
to create a DNA-based database for cattle
started in 1997 and we started DNAbased parentage testing in July 2000.
By October 2004, DNA-based parentage
testing completely superseded the blood
typing system and the DNA database is
now fully operational, with all testing
carried out using DNA.”
The cattle DNA database is used for
various objectives like monitoring
registration and parentage testing,
verifying biotechnology techniques such
as artificial fertilization and embryo
transplantation, and for meat traceability.
Meat traceability is one of several recently
implemented government initiatives to
win back consumer confidence following
the discovery of a bovine spongiform
encephalopathy (BSE)-infected cow in
Japan in 2001. All steps in the movement
of beef in Japan are subject to traceability
testing, from birth to slaughter, storage,
The Division of Animal Genetics at the
Maebashi Institute of Animal Science, LIAJ.
and sale at supermarkets, butchers,
and even steakhouses and specialist
restaurants which handle cuts of beef
for final preparation at the diners’ table.
The new traceability legislation has been
actively enforced since December 2004.
Dr Kurogi elaborated: “In Japan, each
animal is identified by a unique 10digit number. The cattle wear ear tags
bearing this number, which is also used
as a key for DNA testing. Each pack of
dressed meat is labeled with the same
10-digit number, and anybody can find
out the past history of each animal from
birth to slaughter using this number.
This information is readily available to
consumers through in-store computers
as well as on the internet through the
National Livestock Breeding Center’s
homepage.”
“The Ministry of Agriculture, Forestry
and Fisheries of Japan has entrusted LIAJ
with the task of storing beef samples
taken after slaughter, as well as individual
identification testing of the stored
samples and samples collected from
retailers. By carrying out the DNA
testing we verify that the traceability
system is functioning precisely as
specified. For this testing, the cattle
DNA analysis technology is essential for
individual identification.”
In response to the recent and rapid
increase in the number of samples
handled for registration and testing, in
2004 Dr Kurogi decided to automate the
DNA extraction process using Tecan’s
Freedom EVO 200 workstation with an
integrated robotic manipulator (RoMa)
arm, 8-channel liquid handling arm (LiHa)
and a PosID™ barcode reader. Dr Kurogi
uses the Freedom EVO with its integrated
Te-MagS™ magnetic bead separation
module to extract and purify DNA from
hair root, blood and tissue samples. “We
operate the Freedom EVO all day, almost
every day. Our extraction program takes
approximately two hours to process one
plate of 96 samples, so it is possible to
handle approximately four plates – 384
samples – in a working day. It is also
possible to operate the Freedom EVO
unattended so, when we need to process
more than four plates
in a day, the workstation is run overnight.
At present, we are extracting DNA from
approximately 75,000 samples in a
year, and the cattle DNA database is
updated daily.”
“We have optimized and standardized
our programs so that we can extract
clean DNA from blood and hair roots
using essentially the same method. It is
very easy to use and all operations can
be carried out by one person, simply by
pressing buttons on a PC. We just line up
the samples in the morning and leave
the workstation to run for the whole
day, only stopping briefly to make slight
adjustments to the program if different
sample types are being processed. We
are now making efforts to shorten the
process time by simplifying the program
without compromising the quality
of results.”
“The Freedom EVO has saved labor and
time by automating the DNA extraction
procedures and the complete barcode
management is extremely useful for us,
eliminating human error. In a testing
facility such as ours, the thoroughness
of sample management is critical so
the PosID barcode identification is
particularly important.”
The flexibility of the Freedom EVO is
allowing the scientists to automate
other laboratory tasks. Dr Kurogi
concluded: “Currently, we are
continuing to transfer the tasks which
were previously performed manually
to Tecan’s workstation. The possibility
to expand the application in response
to increased workload in the future,
of course, is another important
characteristic that attracted us to the
Freedom EVO.”
Dr Kazuhito Kurogi, Dr Tomohito Ito and
Mr Yusuke Kozono with the Freedom EVO
workstation.
The Freedom EVO workstation at the Maebashi
Institute of Animal Science, LIAJ.
23
24
SA M P L E M A NAGEME NT
Managing worldwide
compound delivery with
REMP technology
As a result of the relentless pace of
today’s drug discovery, pharmaceutical
companies face ever-increasing
throughput demands for compound
screening and lead identification.
Many pharmaceutical companies have
active screening libraries that contain
hundreds of thousands or millions of
compounds, and effective and reliable
management of these samples is critical
to drug development processes.
Boehringer Ingelheim is one of
the world’s top 20 pharmaceutical
companies, with headquarters in
Ingelheim, Germany, and 143 affiliated
companies spread around the world.
The company’s principle research site
is based in Biberach, Germany, where
current research focuses include
diseases of the central nervous system,
metabolic disorders and respiratory
diseases. At Biberach, the Screening
Support group relies on several
REMP automated sample storage
and retrieval systems to provide
compound management services for
Boehringer Ingelheim’s high throughput
screening (HTS) laboratories and other
laboratories worldwide.
Dr Gerhard Mihm, head of the Screening
Support group, explained what the
services involve: “On a daily basis, we
need to be able to acquire compounds,
register the compounds into our
corporate and dispensary databases,
store them under appropriate conditions
and/or provide them to our customers
in various HTS and project laboratories,
on time and prepared to high standards
of quality. We deliver compounds in
either solid or dissolved forms and
sample preparation includes weighing,
dissolution and plating. There are only
six of us within the group providing this
service, so it is essential that all of our
processes are highly streamlined and very
efficient, otherwise we could not possibly
cope with these requirements.”
The compounds come from various
sources, including archive compounds
that have been previously synthesized
by Boehringer Ingelheim’s medicinal
chemists, screening compounds from
the combinatorial chemistry groups, and
many externally acquired compounds.
The Screening Support group was
established 15 years ago and its workload
has dramatically increased during this
period. “Over the years, the number of
compounds to be managed has increased
tremendously, as have the numbers of
assays and additional requirements from
our many different customers. In order to
cope with these demands, we rely heavily
on our REMP sample storage systems,”
said Dr Mihm. “We now have more than
800,000 compounds in our collection so
our service would not be possible without
this technology.”
Dosage and Solution Preparation Station
with tube sealing and capping.
The REMP Dosage and Solution
Preparation station.
SA MPLE M ANAGEM E N T
The Screening Support group’s sample store.
“We first started talking to REMP back
in 1996,” continued Dr Mihm, “and
one of the key reasons we chose these
systems was for the rapid service and
maintenance the company could offer
us. We were also convinced that the
technological concepts of the REMP
systems would meet our requirements.
In 1998 we had the first ever REMP
solution store for 96-well tubes installed
and, since then, we have acquired three
more automated REMP systems. These
include stores for our solid samples and
a -20°C store for compound solutions
using REMP 96 and 384 Tube Technology™
consumables and automated cherrypicking functions. Due to the constant
growth of our collection, a new plate
store at -20°C was implemented at the
end of 2006 as well as an expansion of
our vial store to accommodate another
250,000 vials.” With the REMP technology,
the Screening Support group can comply
with Boehringer Ingelheim’s preferred
practice of using single-use only tubes
and plates for the DMSO solutions,
ensuring high quality, contamination-free
samples for all of its laboratories.
In addition to the REMP storage systems,
the group depends on a number of
REMP workstations, including the REMP
Automated Plate Replicator™ (APR) for
copying plates and the REMP Dosage
and Solution Preparation Station™
(DSP; a collaborative development
between Boehringer Ingelheim and
REMP) for automated weighing of solid
compounds in vials with special REMP
weighing caps, and for preparing stock
DMSO solutions and generating 96well mother plates. Compounds that
are already available in microplates or
96-well tubes are dissolved and plated
with a Tecan automated compound
dissolution and reformatting system
(CIPDISS). This new system is based
on a Freedom EVO® liquid handling
workstation and includes a Te-MO™
96 multichannel pipetting option
and a CRS F3 robot. REMP devices are
also integrated with the workstation,
including the Automated Capper/
Decapper™ (ACD96) and the LHS heat
sealer. Boehringer Ingelheim’s engineers
have developed another Freedom
EVO-based workstation (CIVDISS) for
dissolving and plating solid compounds
that cannot be weighed automatically.
“In general, we are happy with REMP
technology. For us, the development of
an excellent relationship with the people
at REMP has been a major benefit.
Communication with them is absolutely
perfect in most cases and it could be
described as more like a partnership
than a vendor-buyer relationship,” Dr
Mihm concluded.
Automated Plate Replicator for
copying plates.
Dr Hans-Peter Sattler, Project Manager, Tecan Integration Group and Nadine
Hauser, Laboratory Assistant, Boehringer Ingelheim.
25
26
C L I N I CA L D I AGNOSTICS
Te-PoolSafe™ boosts
pipetting accuracy
for testing donor blood
The German Red Cross Institute in Frankfurt is responsible for screening thousands of donated
blood samples every day for blood banks in Germany and further afield. Scientists at the Institute
have developed high throughput PCR-based testing methods to screen for a number of viruses,
and samples are pooled prior to screening using three Tecan liquid handling workstations. In
2006, the Institute worked with Tecan to validate the recently launched Te-PoolSafe™ option for
monitoring pipetting during pooling.
Dr Kai Hourfar at the Red Cross Institute.
As one of the first laboratories to use PCR
for testing donated blood, the German
Red Cross Institute in Frankfurt, Germany,
was an ideal candidate for perfecting
Tecan’s new pipette monitoring module
in a rigorous diagnostic setting. Dr Kai
Hourfar, deputy of the Blood Donor
Screening Lab, said: “We demonstrated
years ago that PCR testing of blood
samples saved lives, because we had
cases where some samples were PCRpositive for viruses, even though antibody
tests of the same samples were negative.
Because we were pioneers and still have a
very good reputation for this application,
many institutes from other countries
asked us, and continue to ask us, to
perform PCR testing on their behalf. The
challenge to begin with was that there
was no automation for PCR testing, and
the only feasible way to test the large
numbers of donations was to create mini
pools of the samples. We had worked
with Tecan previously when developing
our serological tests, and decided to
choose Tecan instruments again to
automate pooling for PCR.”
The Institute initially acquired a Genesis
RSP™ liquid handling workstation for
pooling the samples, followed more
recently by two Genesis Freedom®
workstations. All three workstations
are dedicated to pooling the thousands
of samples that the Red Cross Institute
receives each day.
“We create pools of 96 donations,
which means that if we receive 1,000
samples per day then we reduce these
samples to ten pools and only need to
perform ten PCRs for each parameter
being tested,” Dr Hourfar explained.
“We include enrichment procedures in
our method and always use positive
controls with our screens to ensure that
our testing is sufficiently sensitive, and
this approach allows us to meet the
strict testing criteria defined by the Paul
Ehrlich Institute.” The pooled samples
are subjected to PCR and serological
tests simultaneously, as platelets are
only useful for a maximum of around
five days, and this approach clears a unit
of blood for release just one day after
donation. Blood samples arrive at the
Red Cross Institute at night, so pooling
begins at 11:00 pm and is normally
finished by 6:00 am the following
day, ready for the technicians to begin
subsequent procedures. Samples from
first-time donors are always pooled
separately, because these samples have
not been previously screened. However,
the incidence of positive results during
testing at the Red Cross Institute is
extremely low.
CLI N ICAL DIAGNOST I C S
“All the samples are barcoded and
managed by our LIMS software, and we
have several safety measures in place
to prevent any mix-up of the sample
identities,” said Dr Hourfar. “Most
institutes that we test for send us a list
of the samples they are sending and we
compare the tubes we receive with this
list to make sure none are missing or
duplicated. If two samples arrive with the
same barcode, for example, then both
donations will have to be discarded.”
or, alternatively, may be transferred to
the LIMS for automated processing.
Inaccurately dispensed samples are
selectively identified and can be
separately re-pipetted.
Safety precautions are vital in any
diagnostics laboratory and, in 2006, Tecan
launched the Te-PoolSafe option, a liquid
arrival check system based on weight
measurement, that allows blood banks
and nucleic acid testing laboratories to
monitor and evaluate the performance
of their pooling application. The
module ensures full sample traceability
and provides documented proof of
performance, leaving users confident
that the appropriate amount of liquid
has been pipetted from each sample into
the pool. This is a very fast and sensitive
balance with a 16-position tube holder
and it is supported by software that
measures and evaluates each dispense
from primary samples to the sample
pool, to confirm liquid arrival. Results
are displayed to the system operator
“We have pooled well over 5,000,000
donations since we first started testing at
the Red Cross Institute and any additional
safety measures, such as the visual
inspection of archive plates to verify
automated safety features, will always
be important,” Dr Hourfar explained. “It
is critical that we do everything we can
to ensure our pooling procedures are safe
and it is becoming increasingly necessary
in diagnostics laboratories to prove
these safety measures are adequate.
The Te-PoolSafe provides valuable
documentation to show that all blood
pooling steps are accurately performed.”
Dr Hourfar and his colleagues performed
a number of validation studies for the
Te-PoolSafe on behalf of Tecan1, and were
impressed by the module’s potential to
improve monitoring and documentation
during blood pooling.
The Te-PoolSafe option has not been cleared
for use in all countries. Contact your local sales
office for specific information.
Reference:
1. Hourfar MK, Koller M, Roth WK, Kehrli
R, Seifried E, Schmidt M. (2007).
Balance module allows consistent
monitoring and documentation of the
pooling process for NAT-testing. Vox
Sanguinis (in press).
Balancing samples with the Te-PoolSafe.
27
28
EV E N TS 2 0 07
Meet Tecan at these events in the next six months
Americas
American Association for Cancer Research (AACR) Los Angeles, CA 14-18 Apr 2007
Society for Biomolecular Sciences 2007 (SBS)
Montreal
15 Apr 2007
FASEB Experimental Biology Washington, DC 29 Apr - 1 May 2007
American Association for Clinical Chemistry (AACC)
San Diego, CA
15 Jul 2007
Drug Discovery Technology 2007 (DDT)
Boston, MA 06 Aug 2007
The 6th International BioExpo
Tokyo
20 Jun 2007
JAIMA Show 2007
Makuhari
29 Aug 2007
The Japanese Society for the Study of Xenobiotics
Sendai
09 Oct 2007
Sanguin Spring Seminars 2007
Amsterdam
19 Apr 2007
MC-GARD Molecular Profiling of the Genome
Amsterdam
02 May 2007
MipTec 2007
Basel
07 May 2007
The Swedish Microbiology Spring Meeting 2007
Stockholm
09 May 2007
AMT – Advances in Microarray Technology
Edinburgh
15 May 2007
Euromedlab 2007
Amsterdam
03 Jun 2007
European Human Genetics Conference 2007
Nice
15 Jun 2007
ESACT 2007
Dresden
17 Jun 2007
Molecular Life Sciences 2007 – Herbsttagung der GBM
Hamburg
16 Sep 2007
DGTI Jahreskongress
Friedrichshafen
18 Sep 2007
Biotech Forum & Scanlab
Stockholm
24 Sep 2007
ILMAC
Basel
25 Sep 2007
106th General Meeting
Asia and Pacific
The 28th Annual meeting of Molecular Biology Society of Japan
Europe, Middle East and Africa
Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276
Design: OTM/London www.otmcreate.com
Photography: Marc Wetli/Zürich www.wetli.com, Günter Bolzern/Zürich www.bolzern.net,
Susanne Völlm/Zürich www.susannevoellm.ch, Beat Glauser/www.beatglauser.com, Tim Cook/The
Day Publishing Company (New London, CT)
Editor: kdm/UK www.kdm-communications.com
Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch
Address: Tecan Switzerland AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf,
Switzerland, [email protected],www.tecan.com
Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this
publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd.
cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy
or completeness of the information provided in this publication. Changes in this publication can be
made at any time without notice. All mentioned trademarks are protected by law.
For technical details and detailed procedures of the specifications provided in this document
please contact your Tecan representative.
This brochure may contain reference to applications and products which are not available in all
markets. Please check with your local sales representative.
Cellerity, Columbus, Gemini, Genesis RMP, Genesis RWS, GenePaint, HID EVOlution, HS 4800 Pro,
Infinite, MiniPrep, MultiChannel Arm, PosID, ProfiBlot, Safire2, Sunrise, FlaskFlipper, Te-MO,
Te-MagS, Te-PoolSafe, Te-Stack and Te-VacS are trademarks and Freedom EVO, Freedom EVOlyzer,
Freedom EVOware and Genesis Freedom are registered trademarks of Tecan Group Ltd.,
Männedorf, Switzerland.
Tecan is in major countries a registered trademark of Tecan Group Ltd., Männedorf, Switzerland.
Automated Capper/Decapper, Automated Plate Replicator (APR), DSP and REMP Tube Technology
are trademarks of REMP AG, Oberdiessbach, Switzerland.
© 2007 Tecan Trading AG, Switzerland, all rights reserved.
Headquarters:
Tecan Group Ltd., Seestrasse 103, CH-8708 Männedorf, Switzerland
T +41 44 922 88 88 F +41 44 922 88 89 [email protected] www.tecan.com
Tecan
Austria +43 62 46 89 33 Belgium +32 15 42 13 19 China +86 10 586 95 936 Denmark +45 70 23 44 50 France +33 4 72 76 04 80 Germany +49 79 51 94 170
Italy +39 02 215 21 28 Japan +81 44 556 73 11 Netherlands +31 18 34 48 17 4 Portugal +351 21 000 82 16 Singapore +65 644 41 886 Spain +34 93 490 01 74
Sweden +46 31 75 44 000 Switzerland +41 44 922 89 22 UK +44 118 9300 300 USA +1 919 361 5200 ROW +43 62 46 89 33
REMP
Switzerland +41 31 770 70 70 Japan +81 44 542 70 21 USA +1 508 429 2200

Tecan Journal

Transcription

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