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Proceedings Paper

Rapid detection of bacterial contamination in cell or tissue cultures based on Raman spectroscopy
Author(s): Carsten Bolwien; Gerd Sulz; Sebastian Becker; Hagen Thielecke; Heike Mertsching; Steffen Koch
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Paper Abstract

Monitoring the sterility of cell or tissue cultures is an essential task, particularly in the fields of regenerative medicine and tissue engineering when implanting cells into the human body. We present a system based on a commercially available microscope equipped with a microfluidic cell that prepares the particles found in the solution for analysis, a Raman-spectrometer attachment optimized for non-destructive, rapid recording of Raman spectra, and a data acquisition and analysis tool for identification of the particles. In contrast to conventional sterility testing in which samples are incubated over weeks, our system is able to analyze milliliters of supernatant or cell suspension within hours by filtering relevant particles and placing them on a Raman-friendly substrate in the microfluidic cell. Identification of critical particles via microscopic imaging and subsequent image analysis is carried out before micro-Raman analysis of those particles is then carried out with an excitation wavelength of 785 nm. The potential of this setup is demonstrated by results of artificial contamination of samples with a pool of bacteria, fungi, and spores: single-channel spectra of the critical particles are automatically baseline-corrected without using background data and classified via hierarchical cluster analysis, showing great promise for accurate and rapid detection and identification of contaminants.

Paper Details

Date Published: 8 February 2008
PDF: 11 pages
Proc. SPIE 6853, Biomedical Optical Spectroscopy, 68530F (8 February 2008); doi: 10.1117/12.763247
Show Author Affiliations
Carsten Bolwien, Fraunhofer Institute for Physical Measurement Techniques (Germany)
Gerd Sulz, Fraunhofer Institute for Physical Measurement Techniques (Germany)
Sebastian Becker, Fraunhofer Institute for Biomedical Engineering (Germany)
Hagen Thielecke, Fraunhofer Institute for Biomedical Engineering (Germany)
Heike Mertsching, Fraunhofer Institute for Interfacial Engineering and Biotechnology (Germany)
Steffen Koch, Fraunhofer Institute for Interfacial Engineering and Biotechnology (Germany)

Published in SPIE Proceedings Vol. 6853:
Biomedical Optical Spectroscopy
Anita Mahadevan-Jansen; Robert R. Alfano; Alvin Katz; Wolfgang Petrich, Editor(s)

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