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Tecan Journal 

Edition 1 2005 

ISSN 1660-5276 

Cellerity meets customer needs for 

automated cell culture 

page 6 

HS 400 Hybridization Station makes 

light work of genomic DNA patterns 

page 8 

Talk to Tecan’s Customer Support 

page 14

2 GLOBAL NEWS 

Welcome to the 

“new look” Tecan Journal 

Barbara Stobbe and Mark Wang 

Tecan Journal 1/2005 

Thomas Bachmann 

Chief Executive Officer 

(CEO) 

Firstly, we would like to take this 

opportunity to introduce Thomas 

Bachmann, who joined Tecan in February 

2005 as the new CEO. Thomas brings a 

wealth of experience in international 

senior management to his new role and 

will be focusing his efforts on employees, 

customers and investors alike. 

“My strong personal 

commitment to Tecan is 

to continue developing 

excellent relationships 

with our customers and 

partners, and to maintain 

their confidence and 

trust. We will focus on issues that are 

important to our customers 

and aim to win the respect 

of the life science, diagnostic 

and pharmaceutical worlds 

by consistently providing 

the solutions and support 

they need.” 

Training Mark Wang at Tecan Switzerland and Austria 

Our presence in the Asia-Pacific region 

has been boosted by the opening 

of the Tecan office in Beijing. Our new 

Tecan representatives are Mark Wang 

(mark.wang@tecan.com) and Barbara 

Stobbe (barbara.stobbe@tecan.com). 

Distributors in this region recently met at 

the Asia Pacific Distributor Sales Meeting 

2005 in Bangkok. The meeting was 

hosted by Marco Vittoz, Director Sales 

International, and attended by more than 

20 members of Tecan South East Asia 

(SEA) distributors and Tecan representatives 

from China and (SEA), as well as 

new area managers for India and New 

Zealand. During the meeting, information 

was shared between Tecan and the 

distributors on a personal basis and 

Tecan’s sales and support strategy in the 

Asia Pacific market was discussed. 

Asia Pacific Distributor Sales Meeting 2005 in Bangkok

The Tecan OEM components division in 

California’s Silicon Valley has been 

manufacturing OEM instrument 

components under the Cavro® brand for 

more than 30 years. We work directly 

with other companies, giving them 

access to our products and collaborating 

with R&D, sales and service organizations 

to develop and commercialize validated, 

automated solutions. 

Many leading laboratory instrument 

manufacturers already place their trust in 

Tecan’s Cavro OEM components for the 

critical functions of precision liquid 

handling and robotics and, as well as 

relying on our product performance and 

reliability, they also benefit from Tecan’s 

ISO 13485 quality system and QSR 

compliance. 

The RSP 9000 is an XYZ robotic module. 

It is the perfect base unit to automate 

OEM liquid handling applications 

Detection 

Taking microarrays by storm 

Tecan teams up with Novartis to create 

an impressive new microarray technology 

with the potential for use in novel 

applications page 4 

Application Biopharma 

Cellerity TM meets customer needs for 

automated cell culture 

Cellerity, Tecan’s new fully automated 

system for cell line growth and 

maintenance, promises to be the most 

affordable, compact and flexible 

instrument of its kind page 6 

Detection/Application Biopharma 

HS 400TM Hybridization Station makes 


The HS 400 is used to investigate 

molecular changes along different 

pathways in hereditary non-polyposis 

colorectal cancer page 8 

Tecan prepares launch of new hybridization 

stations 

HS 400 Pro and HS 4800 Pro further 

enhance quality page 9 

CONTENTS 

Application Biopharma 

New developments for automation in 

proteomic research 

A robust sample preparation protocol for 

MALDI-TOF MS analysis of tryptic peptide 

extracts using the Tecan TeMO-96 

pipetting robot page 10 

Detection 

GENios Pro TM – the multi-functional 

reader proves a hit for multiplexing 

Tecan’s multi-functional microplate reader 

is used to multiplex HeLa cell-based 

viability and apoptosis assays page 12 

“Tecan’s good customer service is very 

important to us” 

A long-standing customer backs Tecan’s 

continuing commitment to customer 

service page 14 

Events 

Calendar Come and meet Tecan at 

conferences and meetings around the 

world in 2005 

page 16 


3

4 GLOBAL DETECTION NEWS 

Taking microarrays by storm 

A recent collaboration between Tecan and 

Novartis Pharma AG in Basel has produced a new 

highly sensitive microarray technology that has 

the potential to open up new applications and 

capabilities for the pharmaceutical industry. 

Developed by the Custom Microarray 

Laboratory at Novartis, Evanescent 

Resonator (ER) technology produces a 

signal from fluorescence-based 

microarray analyses that is up to a 

hundred times greater than that from 

traditional, passive glass slides. This means 

that less sample material is required to 

give a strong signal, and there is a better 

signal-to-noise ratio so that even 

samples with extremely small 

concentrations can be analyzed. The 

secret of this heightened sensitivity lies 

in the optical configuration of the 

microarray platform, the NovaChip. 

The surface area built into the 

submicrometer region produces an 

interference effect for incident light 

and has the effect of inducing 

stronger resonance. At the same time, 

the excitation energy is restricted to the 

ultra-thin dielectric layer and this is the 


principle behind the evanescent 

resonator. The restriction of the (laser) 

energy to the thin surface layer 

generates enormous electromagnetic 

force. These so-called evanescent fields 

strengthen the fluorescence signal of 

molecules bonded to the surface and 

create a clear demarcation between that 

area and the background fluorescence. 

However, to take advantage of the 

fluorescence strengthening effect of the 

ER principle, it is necessary to adjust the 

angle of incidence of the excitation beam. 

The instruments in the LS Laser Scanner 

series from Tecan are the only commercial 

microarray scanners available that offer 

this option and, just as importantly, give 

highly reproducible results with 

maximum precision*. The LS 200 has 

been in daily use at Novartis since 2002 

and was chosen, not only for the 

LS Reloaded & Connect for microarray 

batch scanning 

adjustable angle, but also because of the 

variety of measuring modes it offers. 

A particular advantage of the new 

technology is the ability to check the 

application of the probe sample, whether 

DNA, oligonucleotides or proteins etc., 

to the slides. Even if the application 

procedure is of the highest quality, with 

such a large number of spots there will 

inevitably be some left without probe 

sample and it is important to register 

these positions before the actual 

experiment in order to avoid false 

negative results. With the LS 200 in 

scatter mode, the buffer residues which 

form on each spot when the probe fluid 

has been successfully applied can be 

detected as salt crystals without 

compromising the array. The LS 200 is 

also completely compatible with a closed 

stacking device, Connect and so this 

checking stage can easily be carried out 

overnight, processing batches of up to 

200 slides, four at a time. 

To take full advantage of the NovaChip 

technology, the quality and 

reproducibility of hybridization is also 

crucial and for this the Novartis team 

also turned to Tecan. Tecan’s HS 4800 

Hybridization Station is completely

HS 4800 for 48 slides processing 

automatic, from pre-hybridization to 

drying the slides and, as there is no 

manual intervention of any sort, it is 

practically impossible for the array to 

become contaminated. The small sample 

volumes and active mixing of the sample 

during hybridization also help to achieve 

maximum sensitivity. There is currently 

no comparable hybridization instrument 

which offers all these features with the 

additional benefit of flexibility of 

throughput for up to 48 slides per run. 

For the Novartis program, the HS 4800 

is set up in the afternoon with up to 

24 microarray slides loaded with the 

fluorescence-marked samples and left to 

run overnight. Integrated automated 

drying of slides with nitrogen protects 

fluorescence labels from oxidization until 

they are measured next morning using 

LS 200 with Connect stacker. 

The data are then automatically analyzed 

using Array-Pro® Analyzer Software from 

Media Cybernetics based in Silver Spring, 

MD, USA. This software takes away any 

chance of operator variability by means 

of the patented fixing of the spot 

localization to the exact pixel and 

automation of the computer algorithms. 

More spots and genes are detectable by ER scanning 

DETECTION 

The calculation is carried out within 

seconds and is therefore ideally suited 

to high throughput, fully automated 

methods such as this. 

The new technology has proven to be 

extremely flexible and can be used for 

any kind of single or dual color array 

application, with standard 3 x 1 inch 

microarray glass slides as well as other 

glass dimensions and chip formats. In 

summary, the NovaChip provides an 

excellent solution for extending the 

applications of microarray technology 

and, at the same time, reduces the 

number of steps in sample preparation. 

*The LS Reloaded, available since late 2004, has 

three times the sensitivity at 532 nm excitation 

wavelength and twice the sensitivity at 633 nm 

excitation wavelength of the LS 200 used by Novartis. 

Signal to background ratio on NovaChip depends on excitation angle 

5

6 APPLICATION BIOPHARMA 

Cellerity TM meets customer needs 

for automated cell culture 

The launch of 

Cellerity TM , Tecan’s 

dedicated system 

for automated cell 

culture, was met 

with resounding 

enthusiasm at the 

Lab Automation 

2005 show in San 

José, California, this 

past February. 

Delegates visiting the Tecan stand and 

guests at a specially held seminar were 

thrilled to see Cellerity in action and all 

signs show that it promises to be the 

most affordable, compact and flexible 

instrument on the market for cell culture 

and maintenance. The new system can 

be relied upon to automate all standard 

cell culture processes such as passaging, 

splitting, counting and plating. It can 

simultaneously grow different cell lines 

and maintain them in parallel. 

Cellerity is the latest product to result 

from Tecan working together with 


The pierceable cap of the Corning RoboFlask vessel helps maintain a sealed environment during 

media exchange. It can be removed for manual access. 

partner companies who are leaders 

in their own particular fields. In the 

development of Cellerity and its 

corresponding consumables, Tecan, 

Corning® and Invitrogen GIBCO® have 

directly responded to the needs of 

laboratories faced with the constant 

demand for a reliable, timely and 

consistent supply of high-quality cells. 

Cellerity is based on the Freedom EVO® 

200 platform with an eight-tip liquid 

handling arm (LiHa) which has four 

channels for handling cells and four for 

adding sterile liquids such as media, 

additives or buffers. An integrated robotic 

manipulator (RoMa) arm transports 

plates and flasks around the workspace. 

The RoMa directly accesses all pipetting 

stations, incubators, storage devices and 

other modules. The system can handle 

automation-friendly cell culture flasks. All 

pipetting and cell manipulation steps are 

performed within a laminar flow sterile 

hood or optionally a class II biosafety 

cabinet. The robotic incubator controls 

CO2, temperature and relative humidity 

and can hold up to 1000 of both flasks 

and assay plates. CellGEM, the new 

Cellerity control software for Cell Growth,

Expansion and Maintenance, is an 

intuitive software interface that guides 

users through the process of initiating a 

cell line expansion request. A calendarstyle 

interface allows easy selection of 

the dates upon which cells are required 

for delivery. The CellGEM scheduling 

software calculates the optimal times for 

splitting, harvesting, and plating and 

then instructs the instrument to perform 

those actions automatically. The software 

maintains a record of the cells grown in 

the system, including growth rates and 

passage number. Additionally, CellGEM 

keeps track of current volumes of media 

and consumables available in the system 

and warns users of depleting resources. 

The whole system can even be left 

unattended over a weekend with 

instructions to produce cells ready for 

Monday morning, effectively providing 

the laboratory with an extra two working 

days. Bar coding of the flasks means that 

the system knows exactly which protocol 

to apply to each flask. 

Standard T-flasks for cell culture are 

not automation-friendly and require 

expensive industrial equipment to 

manipulate them between stages for 

storage, decapping and incubating. To 

address this problem, Tecan has worked 

closely with the Life Sciences Division of 

Corning Inc. to develop the automationfriendly 

RoboFlask vessel for Cellerity, 

designed to meet standard microplate 

dimensions. These flasks can also be 

handled by other existing Tecan hardware 

and can be stored in instruments 

designed to hold microplates, such as 

incubators, stackers and carousels. 

The RoboFlask vessel has been validated 

for the successful growth of many 

different common and rare cell lines, of 

human and other mammalian origin. 

With a smaller footprint than a regular 

75 cm 2 flask, the RoboFlask vessel has a 

cell growth surface area of 92.6 cm 2 , 

allowing the growth of approximately 

10 7 cells per flask. The flask cap contains a 

septum that can be pierced by the LiHa 

numerous times without compromising 

the flask’s sterility because the cap is 

sterilized with ethanol prior to each 

piercing and otherwise remains closed. 

Each flask is fitted with a vent with a 

large hydrophobic gas permeable 

membrane for aeration and pressure 

exchange. 

A high speed AutoLoader stores empty 

RoboFlask vessels and transfers them 

directly onto a device designed for access 

by liquid handling tips, and for shaking 

and knocking of flasks to harvest cells. 

Harvested cells may be pumped into 

Corning’s ProCulture® spinner flasks, 

where cell suspensions are diluted as 

necessary for plating or seeding of new 

flasks, based on the measurement of an 

Innovatis Cedex cell counter. Cellerity has 

been designed to accommodate GIBCO 

cell culture media and reagents providing 

Cellerity customers with high-quality 

cell culture products efficiently and 

affordably. A large refrigerator unit on 

Cellerity holds up to eight bulk GIBCO 

media bags whose contents can be 

warmed by an on-line heater to the 

appropriate temperature immediately 

before dispensing. The connection 

between Cellerity’s liquid handling 

system and GIBCO disposable media 

bags has been designed to minimize the 

risks of contamination. Liquids such as 

cell dissociation products (eg, Trypsin, 

TrypLE Express) or transfection reagents 

(eg, Lipofectamine) which are used in 

smaller volumes can be kept in cooled or 

heated containers on the Freedom EVO 

deck, where they can be directly accessed 

by the LiHa. 

APPLICATION BIOPHARMA 

Tecan’s fully automated system for cell 

line growth and maintenance will greatly 

benefit cell culture laboratories. The 

system ensures improved quality of cells 

for a wide range of applications and frees 

up laboratory personnel for other 

research activities. 

Gentle shaking of cell culture flasks 

The robot-friendly flasks are compatible with 

many Tecan automation options including the 

RoMa arm on the Freedom EVO and the 

AutoLoader 


7

8 DETECTION/APPLICATION BIOPHARMA 

HS 400 TM Hybridization Station makes 


Hereditary non-polyposis colorectal cancer 

(HNPCC) is a familial syndrome that gives a 

much higher risk of developing colon 

cancer and accounts for 2-5 % of all cases. 

Two different molecular pathways are 

thought to be involved in the development 

of this disease, one being punctual 

alterations of the DNA, and the other, 

chromosomal instabilities in 

microsatellites. Dr Wolfgang Dietmaier and 

colleagues at the Regensburg University 

Clinic in Germany have been using Tecan’s 

HS 400 Hybridization Station to compare 

genome-wide DNA profiles in samples 

from these two variants. It has already 

been shown that microsatellite-stable 

(MSS) and microsatellite-instable (MSI) 

tumor variants can be histologically 

differentiated and the aim of this study 

was to compare the frequency distribution 

of individual genes in tumor tissue with 

healthy tissue. 

DNA was extracted from surgically 

removed tumor and normal mucosa 

samples and labelled with Cy3® and Cy5® 

red fluorescence, respectively. It was then 

hybridized against arrays of a defined BAC 

library of healthy samples, consisting of 32 

blocks (4 x 8 blocks), each approximately 

640 spots, dried on slides. This allowed 

direct comparison of the DNA copy 

numbers of the pathological and healthy 

samples, where DNA gains in a defined 

locus in the tumor DNA were represented 

by red spots, and DNA losses were 

represented by green spots. If the tumor 

DNA was present in equal amounts to that 

in the normal mucosa (i.e. a healthy ratio), 

then the two labels neutralized each other 

to an orange colour. 

The HS 400 Hybridization Station made 

the procedure extremely simple. The arrays 


were placed in the special slide holder and 

the carrier was then inserted into the 

instrument in dry form. The entire method 

was carried out in stages – pre-washing, 

insertion of the hybridization sample, 

hybridization, four washing steps and, 

finally, drying with nitrogen – and the 

individual times, temperatures and buffers 

for each stage were optimized for this 

particular application. 

This application highlighted a number of 

advantages that the HS 400 has over other 

hybridization stations, many of which 

concentrate on minimizing the risk of 

contamination and other artifacts at 

different stages throughout hybridization. 

The entirely automated process takes place 

in the unique four microarray slide holder 

including the final drying step, which 

means that the arrays are dry when they 

enter and leave the instrument and do not 

need to be handled throughout the 

hybridization. 

Cy3 and Cy5 are registered trademarks of Amersham, Inc. 

Figure 1: Example of one block of the 

microarray used for profiling of 

hereditary non-polyposis colorectal 

cancer (HNPCC). Automated 

processing in Tecan’s HS 400 leads to 

fewer background artifacts, higher 

sensitivity and specificity compared 

to other procedures.

Tecan prepares launch of 

new hybridization stations 

HS 400 Pro and 

HS 4800 Pro further 

enhance quality, reproducibility 

and application 

flexibility for microarray 

hybridizations 

Tecan’s HS Series of hybridization stations 

are well proven tools to automate a 

variety of microarray applications, including 

DNA and protein arrays, and in situ 

hybridization. 

The new HS 400 Pro and HS 4800 Pro 

contain Tecan’s unique ABS (Active Bubble 

Suppression) system, which prevents 

formation of bubbles within the chambers, 

eliminating hybridization artifacts and 

enhancing consistency of results. Novel 

dual chambers are easily interchangeable 

with single area chambers and enable 

the new systems to fully automate the 

processing of two independent subarrays 

per slide with no risk of carry-over. 

HS 400 Pro TM and HS 4800 Pro TM 

share the benefits of Tecan’s 

existing hybridization systems: 

DETECTION/APPLICATION BIOPHARMA 

• Fully automated hybridization – from 

pre-hybridization to automated slide 

drying 

• Less background, fewer artifacts, 

higher sensitivity 

• Designed to maximize reproducibility 

and reliability of results 

• Easy to use, low maintenance 

• Low volume hybridization chambers 

save precious samples 

• Two models can handle a variety of 

throughputs 

New products 


9

10 APPLICATION BIOPHARMA 

New developments for automation 

in proteomic research: 

MALDI sample preparation 

for high throughput peptide 

mass fingerprinting 

Matrix-Assisted Laser Desorption Ionization 

Time-of-Flight (MALDI-TOF) mass spectrometry 

(MS) is a key technique for the 

analysis of proteins and peptides in proteomic 

research. It can be used to analyze 

molecules over a wide molecular weight 

range from a few hundred daltons up to 

300kDa with very high accuracy and with 

detection sensitivities in the low fmol 

range. It is very useful in peptide mass 

fingerprint analysis of large proteins. 

Sample preparation and the preparation 

of an appropriate analyte/matrix mixture 

are critical limiting factors in MALDI-TOF 

mass spectrometric analysis. To achieve 

high sensitivity, impurities such as salts 

and buffer components must be washed 

away efficiently with minimal loss of 

analytes. 

Figure 1: MALDI sample preparation 

performed on a Tecan Genesis Workstation 

with a TeMO-96 multi-pipetting unit 


The two experiments described below 

evaluate a robust sample preparation 

protocol for MALDI-TOF MS analysis of 

tryptic peptide extracts using the Tecan 

TeMO-96 pipetting robot (Figure 1), which 

aims to make MALDI-TOF more accessible 

for high throughput applications. 

Method 

Our sample preparation is based on the 

α-cyano-4-hydroxycinnamic acid (CHCA) 

affinity preparation method (Gobom et 

al. (2001), Anal. Chem. 73, 434-438) in 

which the tryptic peptide extracts are 

applied onto a pre-structured anchor 

MALDI sample plate (Scout-MTP 384/600 

AnchorChip; Bruker Daltonics, Bremen, 

Germany) prepared with a thin layer of 

the MALDI matrix CHCA. The TeMO 

worktable accommodates eight 96-well 

microtiter plates with peptide samples 

and two MALDI sample plates, each with 

384 sample positions. The TeMO-96 

pipetting head transfers 1 µL of each 

peptide sample from the microtiter 

plates to the MALDI sample plate where 

the droplets are allowed to dry. New racks 

of disposable pipette tips are delivered by 

the Tecan TeStack for each set of 96 

tryptic extracts to avoid sample cross 

contamination. After the tryptic digest 

samples have dried, contaminants are 

washed away by dispensing 3 µL droplets 

of 0.1 % trifluoroacetic acid (TFA) onto the 

sample, followed by aspiration of the 

droplet after three seconds. Between 

washing of different sample spots, the 

pipette tips were rinsed using the Tecan 

washing station in a multi-step 

procedure included in the MALDI sample 

washing procedure. After emptying the 

tips, two cleaner steps with 90 µL MilliQ 

water each were employed, with soak 

Dr. Niklas Gustavsson*, Dr. Dieter Weichart, 

Dr. Johan Gobom, Max Planck Institute for 

Molecular Genetics, Berlin 

*now based at the Mass Spectrometry group, 

Dept of Plant Biochemistry, Lund University, Sweden 

times of 1 second and flow through set at 

80. The first step included three cycles, 

with change of 5 µL each, and the second 

step included two cycles without change. 

Controlling carry-over during MALDI 

sample plate preparation 

The consumption of pipette tips during 

washing of sample spots on the MALDI 

sample plates can be minimized by using 

the same set of tips for washing of all 

samples on an entire sample plate, as 

long as the pipette tips can be rinsed 

sufficiently between washing of different 

sample spots on the MALDI sample plate. 

The efficiency of the pipette tip washing 

procedure was tested by preparing a set 

of three single peptides on adjacent 

sample spots on the MALDI sample plate 

(Figure 2), followed by washing the whole 

sample plate with one set of pipette tips. 

Angiotensin I, neurotensin and ACTH 18- 

39 were added onto quadrant Q1, Q2 and 

Q3 respectively. Nothing was added to 

quadrant Q4. The four quadrants were 

washed using the previously mentioned 

protocol with two cycles of 3 µL of 0.1 % 

TFA each. As shown in the MALDI-TOF 

mass spectra in Figure 2, no carry-over of 

the peptide prepared on the previous 

sample spot could be detected on the 

following spot. None of the peptides 

prepared on quadrants Q1 to Q3 were 

detected in the mass spectrum from 

quadrant Q4. The preparation of the 

three single peptides was performed in 32 

parallel sets, with no carry-over detected 

in any of the mass spectra. 

Assessing reproducibility of 

preparation for MALDI-TOF MS 

The reproducibility of the automatic 

sample application and MALDI sample

plate washing procedures was tested by 

preparing the same set of 368 tryptic 

digests of human proteins excised from a 

two-dimensional gel onto two MALDI 

sample plates. The plates were processed 

in parallel, and spectra were acquired 

under identical conditions on an Ultraflex 

LIFT mass spectrometer (Bruker Daltonics, 

Bremen, Germany). Using the MASCOT 

software, the peptide mass fingerprint 

data were submitted to a database 

search in the Swiss protein database 

(http://www.expasy.org/sprot/), 

considering scores larger than 62 

indicative of reliable identification 

(p

12 DETECTION 

GENios Pro TM 

– the multi-functional reader 

proves a hit for multiplexing 

Multiplexing assays for any applications offer a range of benefits over 

standard assays. In addition to being amenable to high throughput, they 

also allow the measurement and analysis of more than one parameter 

simultaneously and save time and money by conserving precious compounds 

and cell culture reagents. 

However, for multiplexing to really work, 

there is the challenge of developing 

homogenous assays that do not require 

washing and sample handling steps, 

and the requirement of excellent 

performance, flexibility and multifunctionality 

from the detection 

instrument used. 

Prof. Dr. Gadek-Wesierski and her research 

team at the Medical University of Vienna 

have shown that the GENios Pro multifunctional 

microplate reader from Tecan 

meets all these criteria and has a number 

of features such as individual optics 

for all reading modes that make it 

particularly suitable for multiplexing of 

cell-based assays with various readouts . 

The team in Vienna aimed to multiplex 

two cell-based assays having fluorescent 


and luminescent output to obtain 

information regarding viability and 

apoptosis directly from the same sample 

well, using Cisplatin (CP) as a test 

substance for treating HeLa cells. CP is a 

widespread anticancer drug used for 

chemotherapy. It inhibits viability and 

proliferation and induces apoptosis in 

human cervical carcinoma cells (see 

Figure 1). 

In the experiment, HeLa cells S3 were 

cultured in 96-well plates and treated 

with increasing concentrations of CP, the 

CellTiter-Blue® Cell Viability Assay from 

Promega was used to calculate viability. 

This assay determines the number of 

metabolically active cells based on the 

direct addition of resazurin to cells in a 

culture medium, which is then converted 

into resorufin by the cells. Measurements 

Dr. Gerlinde Zerza-Schnitzhofer, 

Dr. Manfred Lansing, Mag. Marieta 

Gueorguieva 1 , Mag. Carmen Ranftler 

and A.o.Univ. Prof. Dr. Jozefa Gadek- 

Wesierski 1 

Tecan Austria GmbH, Grödig, Austria and 

1 Medical University of Vienna, Vienna, Austria 

Figure 1: Detection of apoptotic HeLa cells. 

Untreated HeLa cells and HeLa cells treated with 

40 µM cisplatin (15 hours) were fixed with 

methanol and stained with a fluoresceinconjugated 

monoclonal mouse antibody 

(M30-CytoDeath, Roche, Vienna, Austria). The 

CytoDeath antibody is specifically for caspase-3 

cleaved cytokeratin-18 and stains cells in various 

stages of apoptosis. Cell nuclei were made 

visible using DAPI. Mitotic (M) and apoptotic (A) 

cells are marked with the following symbol ->.

were taken after the appropriate 

incubation period using the GENios Pro 

reader (Figure 2). Resorufin is activated at 

545 nm and emission is detected at 

590 nm. The fluorescence reading can 

then be correlated with the number of 

metabolically active, living cells, if 

appropriate serial dilutions of cells have 

been performed accordingly. 

In experiments with adherent cells, 

excitation and detection through the 

bottom of a microplate are preferable to 

top reading as this reduces disruptive 

influences of the culture medium on the 

measurement signal. A preliminary test 

with CellTiter-Blue® showed that the 

detection limits in bottom reading mode 

were between six and eight times lower 

than in top reading mode. This means 

that fewer cells and materials can be 

used to achieve the same results, which is 

particularly advantageous for cell-based 

applications. 

Figure 2: GENios Pro microplate reader. Features 

such as rapid fluorescence bottom reading and 

reagent addition by injectors make this 

multimode reader particularly suitable for a 

large number of cell-based assays in HTS. 

Figure 3 shows the results of the CellTiter- 

Blue® assay after the HeLa cells were 

incubated for 24 hours with various 

concentrations of CP (0, 1, 5, 20, 40 µM). 

As the CP concentration increases, the 

number of metabolically active cells is 

reduced by approximately 70 %. 

Information on the apoptosis rate was 

obtained in the multiplex assay using the 

Promega Caspase-Glo 3/7 assay to 

measure the activity of caspase-3 and -7, 

both of which play a key role in the 

apoptosis of mammalian cells. Caspase 

activity is measured by adding a 

preluminescent substrate solution into 

Prof. Dr. Gadek-Wesierski and her 

research team are based at the 

Institute of Cancer Research at the 

Medical University of Vienna. The 

institute is focused on basic cancer 

research, scientific education and the 

development of a new understanding 

of cancer disease. 

The group is studying the mechanisms 

of cell cycle regulation during 

malignant transformation of cells. The 

expression, activation and inhibition of 

tumor suppression proteins (eg, p53), 

Figure 3: Multiplex analysis of viability and 

apoptosis in HeLa cells. Viability was analyzed 

using CellTiter-Blue® Assay and apoptosis using 

Caspase-Glo 3/7 Assay following incubation of 

the cells in 96-well microplates with 0 – 40 µM 

cisplatin (CP). 

the sample wells containing the CellTiter- 

Blue reagents, after having taken the 

viability readings. The substrate solution 

contains a stabilized luciferase and is 

optimized for caspase/luciferase activity 

and cell lysis. The substrate solution 

destroys the cells, the preluminescent 

substrate is cleaved by active caspases 

releasing luciferin that is then consumed 

by the luciferase. The resulting 

luminescent signal is proportional to 

caspase activity and can be measured. 

The results in Figure 3 show a clear 

increase in apoptosis at high CP 

concentrations. The number of apoptotic 

cells increased in this experiment by 

approximately four fold. Correlating the 

readings from both the viability and 

apoptosis analyses with the number of 

cells allows the percentage of apoptotic 

cells to be determined. 

Summary 

Multiplexing is an efficient way of 

obtaining more than one set of 

experimental data from the same sample 

well in order to better understand 

CUSTOMER DETECTION 

SUPPORT 

which play a key role in cycle control, are 

investigated in different cell lines and 

species. Additionally, the group is studying 

the effect of new anti-cancer drugs, in 

particular regarding their potential for 

reactivating the tumor suppressor protein 

p53, which results in enhanced efficiency 

of chemotherapy treatment. 

http://www.p53parp.at 

complex cellular processes. Additionally, 

multiplexing offers the advantage of 

reducing experimental variation, like 

fluctuations in cell numbers due to cell 

clumping and dispensing. Homogenous 

assays, which require no washing and 

separation stages, can easily be 

integrated into Tecan Robotic systems 

with the GENios Pro Reader for use in 

high throughput screening. Stages such 

as the addition of various assay reagents 

by injectors into 96 and 384 well plates, 

incubation at 37° C and plate-shaking, 

can all be carried out in this modular 

reader. The measurement methods 

available on the GENios Pro are suitable 

for a large number of multiplex assays 

and range from absorption and 

fluorescence (FI, FRET, TRF) to various 

luminescence techniques (BRET2, Glow 

and Flash Type Luminescence). 

This experiment combined two cell-based 

assays in one multiplex experiment but, 

using additional assays, e.g. LDH release 

or reporter gene assays, the team 

believes that multiplexing could easily be 

extended to answer further questions. 

Acknowledgement 

We would like to thank Prof. Dr. Gadek- 

Wesierski and her team at the University 

of Vienna for providing the data and 

Dr. Katarina Bohm and Dr. Heinz Winkler 

from Promega for kindly providing the 

reagents. 

GENios Pro is a trademark of Tecan Group Ltd. 

Cell Titer-Blue® and Caspase-Glo 3/7 Assays are trademarks 

of Promega corporation. 

BRET2 is a proprietary technology from BioSignal Packard. 

Patents pending. 


13

14 CUSTOMER SUPPORT 

Tecan Journal 2005 

Just a 

friendly 

phone 

call away

Laurent Nussbaumer is Supporting Engineer at 

Syngenta’s Crop Protection Research facilities 

based in Basel and Stein in Switzerland. 

Mr Nussbaumer is responsible for equipment at 

both sites and has worked with Tecan on several 

projects over a period of seven years. 

What is the main role of your research 

facilities? 

As a whole, the Crop Protection Research 

Facilities are responsible for investigating 

chemicals that have the potential to 

protect crops, mainly fungicides, 

insecticides and herbicides. The site in 

Basel is essentially a dispensary, where all 

the research compounds are stored, 

prepared into solution and pipetted by 

Tecan systems into reaction microplates. 

These plates are then transferred to the 

facility in Stein where we perform a 

range of preliminary and validatory tests, 

also using Tecan systems, on the 

chemicals themselves, and on the effects 

they have on plants in different 

formulations and concentrations. 

What level of sample throughput are you 

achieving? 

At the moment our throughput is around 

200-300 compounds per week, not what 

I would call high throughput at all, but 

we are performing numerous tests at 

many different concentrations with each 

individual compound. 

What Tecan equipment do you have and 

how long have you been using it? 

We have been using Tecan equipment 

since we started operations seven years 

ago. At the Basel site, we have two 

GENESIS RSPs with liquid handling (LiHa) 

arms on each for pipetting. A Freedom 

EVO and carousel handle the plates so 

the process is completely automated. In 

Stein we have GENESIS RSPs equipped 

with LiHa and robotic manipulator 

(RoMa) arms, carousels and specially 

adapted needles that spray the 

chemicals onto the plants. 

Did Tecan provide you with application 

support to set up these processes? 

Yes, we have had application support 

from the beginning. At first, our 

equipment was set up for high 

throughput screening but every time we 

have a new project or change the process 

we have needed to adapt the equipment. 

Sometimes we can make the changes 

ourselves, but if we need instrument 

upgrades or other alterations we always 

directly involve Tecan. 

And how do you rate Tecan’s customer 

service? 

Very good! First and foremost, the 

equipment is generally very reliable. 

However, whenever there is a problem, the 

Tecan engineers guide us through some 

general checks so that they can fully 

analyse the problem and advise whether 

there is some way we can resolve the 

problem ourselves. If not, they are with us 

within 24 hours or if possible, because we 

are quite close, within 3 hours. Tecan’s good 

customer service is very important to us. 

CUSTOMER SUPPORT 

Tecan Customer 

Support 

Whether the customer needs 

assistance in reliable automation, 

precise and safe liquid handling, or 

maintenance and operation of robotics 

and detection products, services are 

in place to assist the customer at all 

levels of activity. In this way, Tecan is 

able to offer the expertise necessary 

to ensure professional use and 

high-performance operation of 

Tecan equipment. 

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 

Editor: kdm/UK www.kdm-communications.com 

Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch 

Address: Tecan Switzerland AG, Marketing Communication, Seestrasse 103, CH-8708 Männedorf, journal@tecan.com, www.tecan.com 

©2005, Tecan Trading AG, Switzerland, all rights reserved 


15

16 GLOBAL NEWS 

Conferences and trade shows 2005 preview 

Europe 

SBS Geneva September 11-15 2005 

Biotech Forum and Scanlab Stockholm October 10-12 2005 

Biotechnica Hannover October 18-20 2005 

JIB – Journées Internationales de Biologie Paris November 3-5 2005 

International Biotech and Lab Automation Europe London November 15-16 2005 

Medica Düsseldorf November 16-19 2005 

Expoquimia Barcelona November 14-18 2005 

Japan 

The 5th Protein Science Society of Japan Annual Meeting Fukuoka June 30-July 2 2005 

The Japan Society for Clinical Laboratory Automation Yokohama September 28-30 2005 

The 28th Annual Meeting of Molecular Biology Society of Japan Fukuoka December 7-10 2005 

USA 

AACC Orlando, FL July 24-28 2005 

Drug Discovery Boston, MA August 7-12 2005 

Chips to Hits Boston, MA September 12-15 2005 

International Symposium on Human Identification (Promega) Grapevine, Texas September 26-29 2005 

Neuroscience Washington, DC November 12-16 2005 

ASHG Salt Lake City, Utah October 26-29 2005 

AABB Seattle, WA October 16-18 2005 

Cell Biology (ASCB) San Francisco December 10-14 2005 

Australia 

2. ComBIO 2005 Adelaide September 25-29 2005 

Asian Pacific Congress of Clinical Biochemistry Brisbane September 14-18 2005 

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 42 334 88 55 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 Other +43 62 46 89 33 

Tecan Journal 1/2005 www.tecan.com

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