www.gbo.com/bioscience

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www.gbo.com/bioscience

Biochips

Technical Information 10 I 2

HTATMPlatforms 10 I 4HTATMSlides 10 I 4HTATMPlate 10 I 6HTATM Buffer Systems 10 I 7CheckScannerTM 10 I 8CheckReportTMSoftware 10 I 8

Diagnostic Kits 10 I 9PapilloCheck® 10 I 9PelvoCheck® 10 l 9ParoCheck® 10 I 10CarnoCheck® 10 I 10

Mycoplasma Detection 10 I 11CytoCheck® 10 I 11Mycoplasma Service 10 l 11

Microfluidics

Customised Microfluidic Platforms 10 I 12

10 Biochips / Microfluidics

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HTATM High-Throughput microArraying

Introduction

The complete deciphering of the human genome and the rapidly growing number of completely sequenced animal, plant and microbial genomes open up new investigative possibilities and techniques. Complex functional analyses, mutation detection or genotyping can be performed within a few hours with the aid of microarrays or „biochips”.Greiner Bio-One develops and produces high-quality biochips for human diagnostics, consumer protection and quality control in the pharmaceutical industry. Our expertise encompasses the development and production of our own innovative biochip platforms for the parallel analysis of samples, the selection of probes, the validation and production of biochips as well as their certification as In-Vitro Diagnostic Devices according to the European Guideline 98/79/EC.

HTATMPlatforms

In academic research with “high-density microarrays”, some samples are investigated for thousands of parameters whilst compared to diagnostics thousands of samples are analysed for a few expressive markers. Therefore parallel comparison and automation play a decisive role. In order to take this into account, Greiner Bio-One has developed new HTS platforms made from plastic especially for use in biochip technology – the HTATMPlatforms (High-Throughput microArraying).

•HTATMSlide1: slide with even surface

The following low-autofluorescence HTATMPlatforms are available:

•HTATMPlate: plate with 96 wells

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• HTATMSlide12: slide with 12 wells

All HTATMPlatforms are barcode-labelled on request.!

Surface

multifunctional (MF)

3-D Amino (AM)

3-D Aldehyde (AL)

3-D Epoxy (EP)

++ most suitable+ suitableo not recommended

Application

• oligonucleotides• PCR fragments

+

++

+

+

• proteins• peptides

+

+

++

++

• modified oligonucleotides

+

o

++

++

HTATM Surfaces

up to 50 nm

0

Reactive, specific functionality

Unreactive moiety results in very low non-specific binding

Spacer

Substrate

Porous, swellable3-dimensionalfunctional matrix

High stability of the function-defining layer:• Long shelf life• High chemical / mechanical robustness

Exact material design through fine tuning of:• Substrate• Morphology• Chemical functions

Beside the multifunctional surface, Greiner Bio-One offers a proprietary 3-D chemistry, which achieves an excellent binding of molecules to the surface via ionic and covalent reaction pathways. The contact angle is about 45°. The advantages of the 3-D matrix include high sensitivity and a chemical and mechanical robustness that permits using contact as well as contact-free printing systems.

A large variety of user-specific applications can be covered with the help of the surfaces offered by Greiner Bio-One. The overview below can serve as an orientation aid for dif-ferent microarray applications.

The high quality of the HTATM 3-D technology ensures an outstandingly uniform spot morphology and avoids variable background fluorescence which can arise due to unspecific binding. The reproducibility and reliability of the analysis results within an array and between arrays represent an important feature of the HTATM technology, which saves sample material, time and money.

In addition, Greiner Bio-One can fulfil customer wishes for 3-D surfaces with different contact angles, thanks to the flexible production procedure (minimum order size required).

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HTATMPlatforms

10 4 www.gbo.com/bioscience

Cat.-No.

Description

Surface functionalisation

Tests per slide

Slide material

Slides per case

445 825

HTATMSlide1

multifunctional

1

plastic

25

446 820

HTATMSlide12

multifunctional

12

plastic

5

445 830

HTATMSlide1

3-D Amino

1

plastic

5

445 835

HTATMSlide1

3-D Amino

1

plastic

25

446 830

HTATMSlide12

3-D Amino

12

plastic

5

HTATMSlides Multifunctional3-D Amino3-D Aldehyde3-D Epoxy

• HTATMSlide12: 36 mm2 printable area/well

• HTATMSlide12: Low well rim of only 0.5 mm

• Low autofluorescence

• Barcode-labelling on request

446 8XX

445 8XX

HTATM Platforms HTATMSlides

HTATMSlide1The plastic slide HTATMSlide1 has the dimensions of a standard 25 x 75 mm glass slide and can be used for the processing of one sample (Fig. 1).

HTATMSlide12The HTATMSlide12 is also a plastic slide and has the same dimensions as the HTATMSlide1 but is partitioned into 12 flat compartments, each with a printable surface of 6 x 6 mm and a low well rim of only 0.5 mm (Fig. 2). 12 samples can be processed simultaneously. Therefore the HTATMSlide12 is able to combine the advantages of the universal microplate platform with microarrays for diagnostic applications due to an identical well geometry compared with a standard 96 well microplate.

Figure 1: Slide geometry of HTATMSlide1

Figure 2: Slide geometry of HTATMSlide12

A = 75 +/- 0.2 mmB = 25 +/- 0.1 mmC = 1.0 +/- 0.05 mmD = 18.5 mmH = Flatness –< 0.05 mm

A = 75 +/- 0.2 mmB = 25 +/- 0.1 mmC = 1.0 +/- 0.05 mmD = 0.5 mmE/F = 9 +/- 0.02 mmG = 3.0 mmH = Flatness –< 0.05 mm

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Free of detectable DNase, RNase

HTATMPlatforms

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Cat.-No.

Description

Surface functionalisation

Tests per slide

Slide material

Slides per case

445 850

HTATMSlide1

3-D Epoxy

1

plastic

5

445 855

HTATMSlide1

3-D Epoxy

1

plastic

25

446 850

HTATMSlide12

3-D Epoxy

12

plastic

5

Cat.-No.

Description

Surface functionalisation

Tests per slide

Slide material

Slides per case

445 840

HTATMSlide1

3-D Aldehyde

1

plastic

5

445 845

HTATMSlide1

3-D Aldehyde

1

plastic

25

446 840

HTATMSlide12

3-D Aldehyde

12

plastic

5

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Cat.-No.

Description

Surface functionalisation

Tests per plate

Material

Quantity per bag/case

791 840

HTATMPlate

3-D Aldehyde

96

plastic

5

791 830

HTATMPlate

3-D Amino

96

plastic

5

791 820

HTATMPlate

multifunctional

96

plastic

5

791 850

HTATMPlate

3-D Epoxy

96

plastic

5

With 96 shallow wells and a low well rim of only 0.3 mm, the HTATMPlate has been optimised for the simultaneous analysis of multiple samples. The low height of the rims facilitates both quick and cost-effective printing of a wide range of analytes into the 96 „wells“ of the plate. Additionally modern robotic spotters and arrayers are already equipped to handle this 96 well format. The partition of the plate into 4 individual strips containing 24 wells each gives the user a high degree of flexibility permitting processing of a variable number of samples each day. For processing, a removable wash collar allows a temporary increase in volume for hybridisation and washing procedures, and is removed prior to detection. The dimensions of the HTATMPlate are compatible with automated systems.

Figure 3: Schematic structure of the HTATMPlate: (1) frame, (2) 24 well strip, (3) 24 well wash collar

Figure 4: Geometry of a HTATMPlate strip with 24 wells

HTATMPlatforms

HTATMPlate Multifunctional 3-D Amino3-D Aldehyde3-D Epoxy

• 96 elevated wells/ stages with a printable area of 36 mm2 each

• 4 individual strips with 24 wells each

• Low autofluorescence

• Low well rim of only 0.3 mm

• Barcode-labelling on request

10 6 www.gbo.com/bioscience

791 8XX

HTATMPlate

A = 81.5 mmB = 26.95 mmC = 6.4 mmD = 9.0 mm

1

2

3

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Cat.-No.

Description

Concentration

Volume [ml]

Quantity per case

445 055

sciSPOT-Protein

2x

25

1

445 052

sciBLOCK

5x

500

1

445 054

sciBIND

2x

1.6

1

445 051

sciWASH-Protein

8x

500

1

Cat.-No.

Description

Concentration

Volume [ml]

Quantity per case

445 016

sciHYB

1x

1.5

1

445 011

sciWASH I

5x

500

1

445 012

sciWASH II

5x

500

1

445 013

sciWASH III

5x

500

1

Cat.-No.

Description

Concentration

Volume [ml]

Quantity per case

445 015

sciSPOT-MF

2x

50

1

445 018

sciSPOT-AM

2x

50

1

445 017

sciSPOT-AL

2x

50

1

445 014

sciPROCESS-MF

2x

500

1

445 020

sciPROCESS-AM

2x

500

1

445 021

sciPROCESS-AL

2x

500

1

Greiner Bio-One and SCIENION have developed integrated and complete systems for the production and use of DNA- and protein microarrays on the HTA™Platform.

Buffer systems for DNA-microarrays Optimised buffers for each of the three surface modifications are available guaranteeing optimal conditions for spotting and processing:• sciSPOT-MF and sciPROCESS-MF for spotting on multifunctional surfaces• sciSPOT-AM and sciPROCESS-AM for spotting on 3-D Amino surfaces

• sciSPOT-AL and sciPROCESS-AL for spotting on 3-D Aldehyde surfacesThe hybridisation buffer sciHYB creates the most favourable environment for hybridisation, while minimising cross-hybridisation. The washing buffers sciWASH I – III reduce the background and deliver a high level of reproducibility.

Buffer systems for protein microarrayssciSPOT-Protein is a unique spotting solution which facilitates a uniform binding of proteins, antibodies and peptides to HTA™Slides. sciBLOCK buffer removes unbound proteins after spotting and blocks the HTA™ surfaces during incubation. sciBIND buffer creates the most favourable environment for binding reactions. Finally, sciWASH-Protein buffer is a microarray wash buffer which is formulated to promote specific binding and to reduce background signals.

HTATMPlatforms

HTATM Buffer Systems

10 7www.gbo.com/bioscience

445 052445 051

445 055

445 054

HTATM Buffer Systems

• Complete and integrated systems for DNA and protein applications

• Optimised buffer systems for each HTA™ surface modification

• Ultra-pure reagents

• Purified by sterile filtration

• Premixed

• Minimised array-to-array variation

• Expedited data analysis

Detailed protocols for each application are available on www.gbo.com/bioscience/manuals.!

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CheckScannerTM

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CheckScanner™ and CheckReport™Software

The CheckScanner™ is a PC-controlled biochip fluorescence scanner for the laboratory use in combination with the CheckReport™Software. It is designed to scan with high performance ready-to-use microarrays in HTA™Slides manufactured by Greiner Bio-One.

The coordinated parallel development of compatible scanner and software permits the fully automated detection and analysis of up to 4 HTA™Slides (Fig. 5).

Through the use of innovative control systems on the individual diagnostic kits from Greiner Bio-One, which are monitored by the CheckReport™Software during the evaluation, false negative or false positive results are virtually excluded.

The CheckReport™Software is now (from version 4.0) available for two different applications:

• The established CheckReport™Software Basic Version has been developed on the basis of several international standards including DIN EN ISO 13485.

• The new CheckReport™Software US Basic Version has been designed to allow FDA compliance in accordance with the terms of the Code of Federal Regulations 21 CFR Part 11: Electronic Records.

Both are composed of three subsystems: • CheckReport™SampleSheet: This unequivocally links patient data with an individual barcode on each biochip.

• CheckReport™Result: The module is based on the successful GeneSpotter software from MicroDiscovery and contains the actual evaluation.

• CheckReport™Admin: This module controls the user management and prevents the misuse or manipulation of data.

The combination of CheckReport™Software with the CheckReport™Software Plug-Ins PapilloCheck®, PelvoCheck®, ParoCheck®, CarnoCheck® and CytoCheck® allows the safe, user-friendly and automatic evaluation of the data.

CheckScannerTM CheckReportTMSoftware

862 070

CheckScannerTM and CheckReportTMSoftware

New: An integrated software interface allows the integration with different Laboratory-Information-Management-Systems (LIMS).

!

Cat.-No.

Description

Quantity per case

862 080

CheckReport™Software

Basic Version

1

862 070

CheckScanner™

1

862 090

CheckReport™Software

US Basic Version

1

New

Cat.-No.

Description

Quantity per case

862 084

CheckReport™SoftwarePlug-In CarnoCheck®

1

New

862 082

CheckReport™SoftwarePlug-In CytoCheck®

1

New

862 083

CheckReport™SoftwarePlug-In ParoCheck®

1

New

862 086

CheckReport™SoftwarePlug-In PelvoCheck®

1

New

862 081

CheckReport™SoftwarePlug-In PapilloCheck®

1

New

Figure 5: Fully automated analysis of 4 HTATMSlides in a CheckScannerTM

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PapilloCheck®, PelvoCheck®, ParoCheck®, CarnoCheck® and CytoCheck® are part of the oCheck® product line from Greiner Bio-One GmbH

10 9www.gbo.com/bioscience

Diagnostic Kits

PapilloCheck® PelvoCheck®

Diagnostic Kits

All of the Greiner Bio-One ready-to-use kits consist of complete test systems with all the necessary buffers and solutions. Extensive on-chip controls ensure error-free results.

PapilloCheck®

High-risk types of human papilloma viruses (HPV) are causative agents in the occurrence of cervical cancer.

The PapilloCheck® CE-IVD diagnostic kit is intended to be used for the qualitative detection and genotyping of 24 types (18 high-risk and 6 low-risk types) of the human papilloma virus in DNA preparations from human cervical smears. The assay is based on the detection of a fragment of the E1 gene of the human papilloma virus and allows the simultaneous processing of 12 cervical specimens.

Prior to the application of the PapilloCheck® diagnostic kit, DNA has to be extracted from a cervical smear specimen previously collected from the patient using the PapilloCheck® Collection Kit. After the following amplification of a 350 bp fragment of the E1 gene via polymerase chain reaction (PCR) the fluorescence-labelled amplification products are hybridised to specific DNA probes fixed on the PapilloCheck® chip. Due to the fluorescence signal the presence of HPV DNA is visualised by the CheckScannerTM and analysed by the corresponding CheckReportTMSoftware.

Nearly error-free results are guaranteed due to the integration of controls for the quality of the DNA extraction, PCR, spot homogeneity and the hybridisation efficiency. In addition, the integration of dUTP in the PapilloCheck® MasterMix ensures the elimination of carry-over contaminations from previous PCR reactions.

PelvoCheck®

Pelvic inflammmatory diseases (PID) are mostly associated with sexually transmitted bacteria causing millions of new infections worldwide per year. Unidentified infections may lead to dramatic symptoms including complications during pregnancy, blindness of the newborn and infertility.

The newly developed diagnostic kit enables the identification of the six most frequently detected bacteria associated with sexually transmitted diseases (Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum). Both cervical and pooled urine samples can be analysed.

The test is based on the detection of a specific DNA fragment of the 16S rRNA Gene using polymerase chain reaction (PCR). The integration of dUTP in the PelvoCheck® MasterMix ensures the elimination of carry-over contaminations from previous PCR reactions.

The scanning procedure, data evaluation and report generation are performed using CheckScannerTM and CheckReportTMSoftware.

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Cat.-No.

Description

Tests per case

465 060

PapilloCheck® (CE-IVD)

identification of 24

human papilloma

viruses

60

465 070

PapilloCheck®

Collection Kit

10

504 060*)

PelvoCheck® (CE-IVD)

identification of 6 bacteria

associated with sexually

transmitted diseases

60

NewNew

*) available 2010

465 060

PapilloCheck® and PelvoCheck® are part of the oCheck® product line from Greiner Bio-One GmbH

Cat.-No.

Description

Tests per case

462 060

CarnoCheck®

identification of 8

animal species

60

460 010

ParoCheck® 10

(CE-IVD)

identification of 10

periodontal pathogens

10

460 020

ParoCheck® 20

(CE-IVD)

identification of 20

periodontal pathogens

10

10 10 www.gbo.com/bioscience

Diagnostic Kits

ParoCheck® CarnoCheck®

All of the Greiner Bio-One ready-to-use kits consist of complete test systems with all the necessary buffers and solutions. Extensive on-chip controls ensure error-free results.

ParoCheck®

Inflammation of the gum and the paradontium are among the most frequent human diseases. Certain bacterial species can serve as indicators to detect such changes early and thus successful treatment is possible. ParoCheck® is approved as an In-vitro Diagnostic (IVD) which can detect up to 20 different characteristic indicative pathogens.

CarnoCheck®

Consumer protection and quality assurance play an increasing role in public awareness, especially in the production of food. Greiner Bio-One has taken into account this need for transparency with the development of the ready-to-use kit CarnoCheck®. With this biochip 8 different animal species (donkey, chicken, horse, turkey, cow, sheep, pig, goat) can be unequivocally identified in food or other products.

For further information, please visit our website www.gbo.com/bioscience.

460 010

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ParoCheck® and CarnoCheck® are part of the oCheck® product line from Greiner Bio-One GmbH

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Mycoplasma Detection Kit

Cat.-No.

Description

Tests

464 060

CytoCheck®

identification of

mycoplasma species

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CytoCheck® Mycoplasma Service

Mycoplasma Detection

464 060

CytoCheck®

In the biopharmaceutical industry, cell cultures are being used increasingly for the manufacture of therapeutic products. Mycoplasma contamination of these cell cultures is a serious problem, as it potentially compromises the safety of the drugs being produced and reduces the final yield. CytoCheck® is a premium test kit for the identification of mycoplasma species in cell cultures and other biological materials. It has been validated in accordance with the guidelines of the European Pharmacopoeia (Ph. Eur. 2.6.7, 2.6.21).

CytoCheck® detects mycoplasma species with a universal mycoplasma probe (including Acholeplasma sp., Spiroplasma sp. and Ureaplasma sp.) and identifies 40 individual species using species-specific probes which can then be used to track the source of contamination. DNA is extracted from the sample and then PCR is performed on the extracted material. The PCR primers amplify conserved and species-specific portions of the “16S-23S rRNA intergenic transcribed spacer” of mycoplasma DNA. Implementation of dUTP prevents carry-over of PCR products, which could become a subsequent source of contamination. The fluorescently labelled amplified DNA fragments are then hybridised to the microarray chip. The chip contains probes for both species-specific targets and a universal probe that detects any species of mycoplasma present in the original sample. Finally, detection of any bound PCR products is achieved with a microarray scanner.

The CytoCheck® DNA-chip has a total of 225 individual measurement points which are used by the CheckScannerTM instrument and subsequently the analysis software, theCheckReportTMSoftware. The integrated platform allows automatic sample tracking, rapid report generation and digital data management (Fig. 6). The CheckReportTMSoftware was especially designed for 21 CFR part 11 conformity to allow FDA compliance of the test procedure. The test can be processed within only 5 hours and sensitivity and specificity are comparable to the cell culture and indicator cell culture methods.

CytoCheck® is offered as a complete ready-to-use kit and includes the CytoCheck® DNA-chips, the PCR MasterMix, buffers and a detailed instruction manual.

Figure 6: The CheckReportTMSoftware automatically generates a report showing the result of the 40 species-specific probes as well as the universal probe

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CytoCheck® is part of the oCheck® product line from Greiner Bio-One GmbH

Mycoplasma Service Greiner Bio-One offer a service for the detection of mycoplasma called Mycoplasma Service!

463 090

Mycoplasma Service

test for detection

of mycoplasma

1

10 12 www.gbo.com/bioscience

The finest structures for the finest ideasDiscover the potential of microfluidics

Through the use of modern manufacturing techniques it is now possible to produce microstructured components for the diagnostics, drug discovery, and research industry made from innovative plastic polymer materials. They offer substantial economical advantages, a large selection of quite diverse materials and surface treatments are available, and large format components can also be economically structured. The principal application areas are:

Analytics/Diagnostics (electrophoresis, lab-on-a-chip)

Chemistry (micromixers, reactors, heat exchangers)

Drug discovery and liquid handling

For the production of microfluidic systems as single-use products, novel and innovative manufacturing techniques are required. A number of manufacturing processes can be utilised and today injection moulding is widely used to produce microstructured parts from plastics. The choice of material depends on the final application of the components. Common selection criteria are:

Transparency

Stability

Chemical resistance

Biocompatibility

Surface properties (e.g. hydrophobicity)

A large number of different polymers are available (e.g. polystyrene, polymethylmethacrylate, polycarbonate, polypropylene). Additional surface treatments after moulding can further improve the properties of the chosen material, and compared to glass, a larger variety of options for surface modifications are available.

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Figure 1: Production of microstructured parts by injection moulding.

Precise manufacturing of the mould insert is critical for the quality of the plastic component. Different methods are used depending on the structural size, precision and aspect ratio of the structure being moulded. Routinely, mechanical micromachining is used to create the required metal moulding tools. Using high-speed tools, microstructures can be produced even on large areas. For extremely precise mould inserts, galvanic/lithographic techniques are available.

Figure 2a: Mould insert made of brass, manufactured by micro-milling.Figure 2b: Detail of a mould insert made of nickel, manufactured by UV-LIGA.

Standard formats allow for fast and cost-efficient production of customer-specific designs from prototyping to production level. A common format in the life science is the microscope slide with a footprint of 25 mm x 75 mm.

Figure 3: Microfluidic Slide. Microchannels and macro-features (reservoirs, through-holes) were produced in one step.

Greiner Bio-One can produce microstructured components in a variety of shapes and dimensions and also larger formats, such as the microplate format, can be structured. In many cases, the microstructured surface must be sealed with a lid in order to produce a closed channel system. Different joining methods, such as laser welding, ultrasonic welding, adhesive techniques and diffusion bonding are used. The incorporation of additional components such as electrodes or membranes into the microstructured part is also possible. To combine different materials, we have special welding and adhesive bonding techniques that have been adapted for microstructures.

Figure 4a: Partly sealed structures for capillary electrophoresis. Figure 4b: SEM image of the cross section through a closed microchannel (100 µm x 50 µm).

10 13www.gbo.com/bioscience

Our serviceWith our team of biologists, chemists, physicists, and engineers, we work closely with our customers. Built on our experience in plastics manufacturing, we can supply an innovative solution, custom-tailored to your requirements.

A large variety of materials is available to realise your design, in addition to which we offer several modification technologies to tailor the surface properties of your microstructured component to optimise it for your application.

We offer our services from the first draft to the product:

Prototyping

Small series production

Mass production

!2a) 2b)

4a) 4b)

Microchannel

Intersection

Reservoir

1000 µm

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