Cell-biological analytics and accredited testing
Fraunhofer Institute for Interfacial Engineering and Biotechnology
Cultivation of cells from primary material (various tissues and species), construction of three-dimensional cell culture systems and tissue engineering are the expertises of the Fraunhofer IGB. We characterize cells both via classical molecular-biological, histological and immuno-histological methods and via flow cytometry. With our experience of 15 years in cell cultivation, we are your partner for complex questions in regenerative medicine, tissue engineering and the development of cell-based assays for toxicology.
The testing of compatibility issues in the development of new materials is an exploratory focus of Fraunhofer IGB. Thereby we focus on the simulation of ex vivo interactions between new or functionalized materials with human tissues to evaluate cytotoxic effects occurring in the production process or during the application as medical device. We offer the biological evaluation of medical devices according to DIN ISO 10993-5: 2009 (Part 5: In vitro-cytotoxicity).
Testing of biocompatibility and cytotoxicity
We perform the examination of biological compatibility of medical devices according to DIN ISO 10993-5 using cell lines. In addition, we offer biocompatibility testing using our patented 3D skin model according to DIN ISO 10993-5 (accredited in-house method).
A further cell-based model, which has been accredited since 2009, is the Caco 2-model for the classification and estimation of oral bioavailability.
Flow cytometry – FACS sorting and characterization
In flow cytometry, through various new antibody-coupled fluorescence dyes, a wide variety of cell populations can be detected and sorted for further purposes of analysis. Areas of application range from blood diagnostics, via cell transfection to the identification of stem and precursor cell populations.
Automated microdissection allows the cutting of the smallest areas out of the matrix-immobilized cell layer. Even individual cell nuclei can be isolated for further molecular-biological analysis.
For biological applications such as the investigation of microorganisms and single cells, the Raman spectrometer can be combined with a light microscope. Our focus is the development of Raman spectroscopic methods for biological applications, e.g. for the non-destructive sterility control or for label-free cell characterization.
Good to know
Automated microdissection allows the cutting of the smallest areas out of the matrix-immobilized cell layer. A microdissection unit consists of a microscope, combined with a laser beam. With this unit, the smallest samples (1-1000 µm) – i.e. individual cells, but also cell nuclei – can be dissected from a tissue section for post-processing. For example tumor cells from the adjacent healthy tissue can be isolated and examined.
A digital video camera files the pictures of a sample to a computer. For the microdissection, the elements marked before at the screen, e.g. certain cell nuclei from a certain tissue or also individual living cells, are dissected from the surrounding tissue with a software-controlled laser beam. Afterwards they are extracted by single laser pulse counter-gravity into a sampling container.
The 355 nm solid laser has an operation accuracy of distinctly less than 1 µm. This procedure guarantees that no unwanted sample elements can get into the container. As the laser reacts for only about a nanosecond during this process, the dissected sample cannot warm up. The procedure is thus totally contact- and contamination-free. Moreover, it ensures the greatest possible protection of the material.
FACS flow cytometry (Fluorescence-activated cell sorting)
The introduction of laser technology has revolutionized classical cell analytics in the past ten years. In flow cytometry, through various new antibody-coupled fluorescence dyes, a wide variety of cell populations can be detected and sorted for further purposes of analysis. Areas of application range from blood diagnostics, via cell transfection for the generation of cell lines and clone isolation to the identification of stem and precursor cell populations from a great variety of tissue types.
Voices from industry | Testing of biocompatibility
"It was particularly important to me that there was an 'individual' contact person, who was involved in the entire correspondence and was familiar with the order and its development. The response upon my request was prompt, the order was completed very reliably in an unexpectedly short time. Such a service is very pleasing! “
ANKATIT-ANKA Guss GmbH
"We thank you very much for the excellent collaboration. You have performed all investigations with highest efficiency and have accompanied our project with all your experience. We can recommend your services anytime without restrictions."
Courage und Khazaka electronic GmbH
FACS analysis of residual leucocytes in blood preparations
“By the introduction of leukocyte depletion in many European countries both the pharmaceutical quality and the general compatibility of the blood components was clearly improved. By the use of so called adhesion filters, as manufactured by MacoPharma amongst others, the number of leukocytes in the preparations was reduced by several orders of magnitude, which has had the consequence that the number of allo-immune reactions has clearly decreased in transfusion medicine. Further, it could be shown that the infection risk by cell resident viruses such as CMV could likewise be lowered. The determination of the residual leukocyte content in the blood components by flow cytometry (FACS) represents a safe and precise control, and corresponds to the current state of the art. Within the scope of a double test series MacoPharma has assigned the execution of FACS measurements to the independent Fraunhofer Institute in Stuttgart. The test series was successfully completed in 2005.”
Dr. Jürgen Zimmermann, MacoPharma, Frankreich
FACS validation of antibody targets for tumor therapy
“Patrys GmbH, a subsidiary of the Australian Patrys Ltd., develops human monoclonal antibodies for cancer therapy; these were identified and isolated by a proprietary pipeline. Most of these antibodies bind at targets, which up to now were not seen in connection with tumor genesis or identified as targets for tumor therapy. The FACS service unit of the Fraunhofer IGB as an independent institution was able to confirm our data and carried out the bonding studies promptly and competently.”
Antonia Hildebrand, Project Manager, Patrys GmbH
- Light microscopy
- Phase contrast microscopy
- Differential interphase contrast
- 3-D imaging, time series
- Fluorescence microscopy
- Fluorescence microscopy with confocal resolution (ApoTome)
- Automated image data processing
- Observation of larger areas by MosaiX scan function, e.g. of cell agglomerates and tissues
- Histology, molecular- and cellbiological as well as biochemical analysis of tissues and media supernatants
- Automated dissection of cell nuclei or matrix fragments from fixed tissue preparations for the molecular-biological analysis (DNA, RNA, protein expression)
- Catapulting of single cells and cell colonies for further cell cultivation (e.g. single cell deposition of transfected cells or purification of primary cell cultures)
- Immunofluorescence measurements of cell surface and intracellular markers
- Single- and multi-color analysis
- Cell cycle analysis
- Apoptosis detection, viability and proliferation testing (e.g. for testing of biocompatibility)
- Cell sorting according to light scatter or fluorescence parameters
- GFP analysis for determination of transfection efficiency or establishment of stable clones
- Hoechst-Efflux for side population analysis
- Antibody binding studies
- Determination of growth factors in cell culture supernatants by Cytometric Bead Array
Establishment of further methods on demand
- Single cell deposition
- Determination of activation antigens (e.g. on platelets for biocompatibility testing)
- Allergy diagnostics (activation of basophilic cells)
- Kinetic measurements (calcium flux)
- Microscopy (AxioVert 200M, Zeiss)
- Micro-dissection (MicroBeam, PALM-Zeiss)
- Laser scanning technology
- Raman spectrometer
- Real-time PCR
- Flow cytometry
Specification flow cytometry
- TurboSort, MakroSort, AccuDrop, CloneCyte, AutoSort
- Water-cooled Coherent Enterprise II Laser (simultaneous operation of 488 nm and UV)
- Air-cooled SpectraPhysics HeNe-Laser (633 nm)
- Digital electronic and QuadraSort with integral software
- External heating/cooling system for better cell viability and color stability
- Data acquisition and processing station PowerMacintosh G4
- FACSDIVA Software 4.0
- Water-cooled laser (488 nm, red diode laser) for four-color analysis
- Data acquisition and processing station PowerMactintosh G4
- CellQuest Pro Software 4.0.2