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Non-disruptive measurement system of cell viability in bioreactors

: Rudek, Florian; Nelsen, B.L.; Baselt, Tobias; Berger, T.; Wiele, M.; Prade, I.; Hartmann, Peter


Farkas, D.L. ; Society of Photo-Optical Instrumentation Engineers -SPIE-, Bellingham/Wash.:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX : 15-17 February 2016, San Francisco, California
Bellingham, WA: SPIE, 2016 (Proceedings of SPIE 9711)
ISBN: 978-1-62841-945-0
Paper 97110Y, 8 S.
Conference "Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues" <9, 2016, San Francisco/Calif.>
Fraunhofer IWS ()
fluorescence; lifetime measurement; saccharomyces cerevisiae; NADH; cell viability; marker less

Nutrient and oxygen transport, as well as the removal of metabolic waste are essential processes to support and maintain viable tissue. Current bioreactor technology used to grow tissue cultures in vitro has a fundamental limit to the thickness of tissues. Based on the low diffusion limit of oxygen a maximum tissue thickness of 200 mu m is possible. The efficiency of those systems is currently under investigation. During the cultivation process of the artificial tissue in bioreactors, which lasts 28 days or longer, there are no possibilities to investigate the viability of cells. This work is designed to determine the influence of a non -disruptive cell viability measuring system on cellular activity. The measuring system uses a natural cellular marker produced during normal metabolic activity. Nicotinamide adenine dinucleotide (NADH) is a coenzyme naturally consumed and produced during cellular metabolic processes and has thoroughly been studied to determine the metabolic state of a cell. Measuring the fluorescence of NADH within the cell represents a non -disruptive marker for cell viability. Since the measurement process is optical in nature, NADH fluorescence also provides a pathway for sampling at different measurement depths within a given tissue sample. The measurement system we are using utilizes a special UV light source, to excite the NADH fluorescence state. However, the high energy potentially alters or harms the cells. To investigate the influence of the excitation signal, the cells were irradiated with a laser operating at a wavelength of 355 nm and examined for cytotoxic effects. The aim of this study was to develop a non-cytotoxic system that is applicable for large-scale operations during drug -tissue interaction testing.