Tissue Culture 1 Cell/ Microplates 2 HTS- 3 Immunology/ HLA 4 Microbiology/ Bacteriology Purpose Beakers 5 Tubes/Multi-
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1 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo- 0 Biochips/ 4 Accessories Cell/
2 0 Biochips / Biochips Technical Information 0 I 2 HTA TM Platforms 0 I 4 HTA TM Slides 0 I 4 HTA TM Plate 0 I 6 HTA TM Buffer Systems 0 I 7 CheckScanner TM 0 I 8 CheckReport TM Software 0 I 8 Diagnostic Kits 0 I 9 PapilloCheck 0 I 9 PelvoCheck 0 l 9 ParoCheck 0 I 0 CarnoCheck 0 I 0 Mycoplasma Detection Kit 0 I CytoCheck 0 I Customised Microfluidic Platforms 0 I 2 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo- Cell/ 0 Biochips/ 4 Accessories
3 HTA TM 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. Cell/ The following low-autofluorescence HTA TM Platforms are available: HTA TM Slide: slide with even surface 0 Biochips/ Cryo- 2 Lids/Sealers/ 3 Reaction Tubes/ HTA TM Platforms 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 especially for use in biochip technology the HTA TM Platforms (High-Throughput microarraying).! All HTA TM Platforms are barcode-labelled on request. HTA TM Slide2: slide with 2 wells HTA TM Plate: plate with 96 wells 4 Accessories 0 2
4 Cell/ HTA TM Surfaces up to 0 nm 0 High stability of the function-defining layer: Long shelf life High chemical / mechanical robustness 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 4. 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 different microarray applications. Reactive, specific functionality Unreactive moiety results in very low non-specific binding Spacer Substrate Exact material design through fine tuning of: Substrate Morphology Chemical functions Porous, swellable 3-dimensional functional matrix The high quality of the HTA TM 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 HTA TM 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). Surface Application multifunctional (MF) 3-D Amino (AM) 3-D Aldehyde (AL) 3-D Epoxy (EP) ++ most suitable + suitable o not recommended oligonucleotides PCR fragments modified oligonucleotides + o proteins peptides Reaction Tubes/ 2 Lids/Sealers/ Cryo- 0 Biochips/ 4 Accessories 0 3
5 Cell/ HTA TM Platforms HTA TM Platforms HTA TM Slides 446 8XX 44 8XX HTA TM Slides Multifunctional 3-D Amino 3-D Aldehyde 3-D Epoxy HTA TM Slide2: 36 mm 2 printable area/well HTA TM Slide2: Low well rim of only 0. mm Low autofluorescence Barcode-labelling on request HTA TM Slide The slide HTA TM Slide has the dimensions of a standard 2 x 7 mm glass slide and can be used for the processing of one sample (Fig. ). HTA TM Slide2 The HTA TM Slide2 is also a slide and has the same dimensions as the HTA TM Slide but is partitioned into 2 flat compartments, each with a printable surface of 6 x 6 mm and a low well rim of only 0. mm (Fig. 2). 2 samples can be processed simultaneously. Therefore the HTA TM Slide2 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 : Slide geometry of HTA TM Slide A = 7 +/- 0.2 mm B = 2 +/- 0. mm C =.0 +/- 0.0 mm D = 8. mm H = Flatness < 0.0 mm A = 7 +/- 0.2 mm B = 2 +/- 0. mm C =.0 +/- 0.0 mm D = 0. mm E/F = 9 +/ mm G = 3.0 mm H = Flatness < 0.0 mm 0 Biochips/ Free of detectable DNase, RNase Figure 2: Slide geometry of HTA TM Slide2 Cryo HTA TM Slide HTA TM Slide2 HTA TM Slide HTA TM Slide HTA TM Slide2 2 Lids/Sealers/ Surface functionalisation Tests per slide Slide material multifunctional multifunctional 2 3-D Amino 3-D Amino 3-D Amino 2 3 Reaction Tubes/ Slides per case Accessories 0 4
6 HTA TM Platforms Cell/ Surface functionalisation HTA TM Slide 3-D Aldehyde HTA TM Slide 3-D Aldehyde HTA TM Slide2 3-D Aldehyde Tests per slide 2 Slide material Slides per case HTA TM Slide HTA TM Slide HTA TM Slide2 Surface functionalisation Tests per slide 3-D Epoxy 3-D Epoxy 3-D Epoxy 2 Slide material Slides per case 2 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo- 0 Biochips/ Accessories
7 Cell/ HTA TM Platforms HTA TM Plate HTA TM Plate Multifunctional 79 8XX 3-D Amino 3-D Aldehyde 3-D Epoxy With 96 shallow wells and a low well rim of only 0.3 mm, the HTA TM Plate 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 HTA TM Plate are compatible with automated systems. 96 elevated wells/ stages with a printable area of 36 mm 2 each 4 individual strips with 24 wells each 3 Low autofluorescence Low well rim of only 0.3 mm Barcode-labelling on request 2 Figure 3: Schematic structure of the HTA TM Plate: () frame, (2) 24 well strip, (3) 24 well wash collar A = 8. mm B = 26.9 mm C = 6.4 mm D = 9.0 mm 0 Biochips/ Figure 4: Geometry of a HTA TM Plate strip with 24 wells Cryo- Free of detectable DNase, RNase 2 Lids/Sealers/ Surface functionalisation HTA TM Plate multifunctional HTA TM Plate 3-D Amino HTA TM Plate 3-D Aldehyde HTA TM Plate 3-D Epoxy 3 Reaction Tubes/ Tests per plate Material Quantity per bag/case Accessories 0 6
8 HTA TM Platforms Cell/ HTA TM Buffer Systems HTA TM Buffer Systems 44 0 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 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.! Detailed protocols for each application are available on 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 surfaces The 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 microarrays scispot-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 scispot-mf scispot-am scispot-al sciprocess-mf sciprocess-am sciprocess-al Concentration Volume [ml] Quantity per case scihyb 44 0 sciwash I sciwash II sciwash III 0 Biochips/ Concentration x x x x Volume [ml] Quantity per case Cryo scispot-protein sciblock scibind 44 0 sciwash-protein 2 Lids/Sealers/ Concentration Volume [ml] Quantity per case 2 x x 00 3 Reaction Tubes/ Accessories
9 CheckScanner TM Cell/ CheckScanner TM and CheckReport TM Software CheckScanner TM CheckReport TM Software 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. ). 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 348. 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 2 CFR Part : 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.! : An integrated software interface allows the integration with different Laboratory-Information- Management-Systems (LIMS). 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. 0 Biochips/ Figure : Fully automated analysis of 4 HTA TM Slides in a CheckScanner TM Cryo CheckScanner CheckReport Software CheckReport Software Basic Version US Basic Version 2 Lids/Sealers/ Quantity per case Reaction Tubes/ Quantity per case CheckReport Software Plug-In PapilloCheck CheckReport Software Plug-In PelvoCheck CheckReport Software Plug-In ParoCheck CheckReport Software Plug-In CarnoCheck CheckReport Software Plug-In CytoCheck 4 Accessories PapilloCheck, PelvoCheck, ParoCheck, CarnoCheck and CytoCheck are part of the ocheck product line from Greiner Bio-One GmbH 0 8
10 Diagnostic Kits Diagnostic Kits Cell/ PapilloCheck PelvoCheck 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 (8 highrisk 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 E gene of the human papilloma virus and allows the simultaneous processing of 2 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 30 bp fragment of the E 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 CheckScanner TM and analysed by the corresponding CheckReport TM Software. 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 6S 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 CheckScanner TM and CheckReport TM Software. 0 Biochips/ s Cryo- Tests per case PapilloCheck (CE-IVD) identification of 24 human papilloma viruses PapilloCheck Collection Kit 0 PapilloCheck and PelvoCheck are part of the ocheck product line from Greiner Bio-One GmbH * ) PelvoCheck (CE-IVD) identification of 6 bacteria associated with sexually transmitted diseases 60 * ) available Lids/Sealers/ 3 Reaction Tubes/ 4 Accessories
11 Cell/ 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 ParoCheck ParoCheck CarnoCheck (CE-IVD) (CE-IVD) identification of 8 Tests per case identification of 0 periodontal pathogens 0 identification of 20 periodontal pathogens 0 animal species 60 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo- 0 Biochips/ 4 Accessories ParoCheck and CarnoCheck are part of the ocheck product line from Greiner Bio-One GmbH 0 0
12 Mycoplasma Detection Kit Mycoplasma Detection Kit Cell/ CytoCheck 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 , 2.6.2). 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 6S- 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 speciesspecific 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 22 individual measurement points which are used by the CheckScanner TM instrument and subsequently the analysis software, the CheckReport TM Software. The integrated platform allows automatic sample tracking, rapid report generation and digital data management (Fig. 6). The CheckReport TM Software was especially designed for 2 CFR part conformity to allow FDA compliance of the test procedure. The test can be processed within only 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. 0 Biochips/ Figure 6: The CheckReport TM Software automatically generates a report showing the result of the 40 species-specific probes as well as the universal probe Cryo- Tests per case CytoCheck identification of mycoplasma species 0 2 Lids/Sealers/ 3 Reaction Tubes/ CytoCheck is part of the ocheck product line from Greiner Bio-One GmbH Accessories
13 The finest structures for the finest ideas Discover 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 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 s. 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. 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo- 0 Biochips/ 4 Accessories Cell/ 0 2
14 Figure : Production of microstructured parts by injection moulding. Precise manufacturing of the mould insert is critical for the quality of the 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. 2a) 2b) 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 2 mm x 7 mm. 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. 4a) 4b) Reservoir Figure 4a: Partly sealed structures for capillary electrophoresis. Figure 4b: SEM image of the cross section through a closed microchannel (00 µm x 0 µm).! Our service With our team of biologists, chemists, physicists, and engineers, we work closely with our customers. Built on our experience in s 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 Microchannel Intersection 000 µm Cryo- Cell/ 0 Biochips/ Figure 3: Microfluidic Slide. Microchannels and macro-features (reservoirs, through-holes) were produced in one step. 2 Lids/Sealers/ 3 Reaction Tubes/ Accessories
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