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11 February 2010 A system for the rapid detection of bacterial contamination in cell-based therapeutica
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Monitoring the sterility of cell or tissue cultures is of major concern, particularly in the fields of regenerative medicine and tissue engineering when implanting cells into the human body. Our sterility-control system is based on a Raman micro-spectrometer and is able to perform fast sterility testing on microliters of liquid samples. In conventional sterility control, samples are incubated for weeks to proliferate the contaminants to concentrations above the detection limit of conventional analysis. By contrast, our system filters particles from the liquid sample. The filter chip fabricated in microsystem technology comprises a silicon nitride membrane with millions of sub-micrometer holes to retain particles of critical sizes and is embedded in a microfluidic cell specially suited for concomitant microscopic observation. After filtration, identification is carried out on the single particle level: image processing detects possible contaminants and prepares them for Raman spectroscopic analysis. A custom-built Raman-spectrometer-attachment coupled to the commercial microscope uses 532nm or 785nm Raman excitation and records spectra up to 3400cm-1. In the final step, the recorded spectrum of a single particle is compared to an extensive library of GMP-relevant organisms, and classification is carried out based on a support vector machine.
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Carsten Bolwien, Christian Erhardt, Gerd Sulz, Hagen Thielecke, Robert Johann, Marieke Pudlas, Heike Mertsching, and Steffen Koch "A system for the rapid detection of bacterial contamination in cell-based therapeutica", Proc. SPIE 7560, Biomedical Vibrational Spectroscopy IV: Advances in Research and Industry, 756009 (11 February 2010);

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