Abstract
A new-type in situ probe has been developed to acquire dark field images of yeast in bioreactors. It has been derived from an in situ bright field microscope that is able to measure cell density in bioreactors during fermentation processes. The illumination part of the probe has been replaced with a dark field device, in which an aspheric condenser is used, so that high contrast dark field images can be obtained. The technique of second imaging is implemented to improve the sharpness of the images by means of a relay lens. This new in situ probe is expected to enable the evaluation of the cell viability without staining owing to modern image processing.
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References
Bittner C, Wehnert G, Scheper T (1998) In situ microscopy for on-line determination of biomass. Biotech Bioeng 60:24–35
Castro-Concha LA, Escobedo RM, Miranda-Ham M (eds) (2006) Measurement of cell viability in in vitro cultures. Plant cell culture protocols (2nd edn). Humana Press, Totowa, New Jersey, The US
Claes JE, Van Impe JF (1999) On-line estimation of the specific growth rate based on viable biomass measurements: experimental validation. Bioproc Biosyst Eng 21:389–395
Heggart H, Margaritis A, Stewart RJ, Pilkington M, Sobezak J, Russell I (2000) Measurement of brewing yeast viability and vitality: a review of methods. Tech q - Master Brew Assoc Am 37:409–430
Huls PG, Nanninga N, Van Spronsen E (1992) A computer-aided measuring system for the characterization of yeast populations combining 2D-image analysis, electronic particle counter, and flow cytometry. Biotechnol Bioeng 39:343–350
Joeris K, Frerichs JG, Konstantinov K, Scheper T (2002) In-situ microscopy: Online process monitoring of mammalian cell cultures. Cytotechnology. 38:129–134
Nattkemper TW, Ritter HJ, Schubert W (2001) A neural classifier enabling high-throughput topological analysis of lymphocytes in tissue sections. IEEE T Inf Technol B 5:138–149
Nattkemper TW, Twellmann T, Ritter H, Schubert W (2003) Human vs. Machine: evaluation of fluorescence micrographs. Comput Biol Med 33:31–43
Pons MN, Vivier H, Remy JF, Dodds JA (1993) Morphological characterization of yeast by image analysis. Biotechnol Bioeng 42:1352–1359
Scheper T (1991) Bioanalytik. Vieweg-Verlag, Braunschweig, Germany
Sonnleitner B, Locher G, Fiechter A (1992) Biomass determination. J Biotechnol 25:5–22
Suhr H, Wehnert G, Schneider K, Bittner C, Scholz T, Geissler P, Jähne B, Scheper T (1995) In situ microscopy for on-line characterization of cell-populations in bioreactors, including cell-concentration measurements by depth from focus. Biotech Bioeng 47:106–116
Thomas CR, Paul GC (1996) Applications of image analysis in cell biology. Curr Opin Biotechnol 7:35–45
Van Imple J, Iserentant D, Vanrolleghem P (1998) Advanced instrumentation, data interpretation and control of biotechnological processes. Kluwer Academic Publishers, Dordrecht, The Netherlands
Vecht-Lifshitz SE, Ison AP (1992) Biotechnological applications of image analysis: present and future prospects. J Biotechnol 23:1–18
Wei N, Flaschel E, Saalbach A, Twellmann T, Nattkemper TW (2005) Reagent-free automatic cell viability determination using neural network based machine vision and dark-field microscopy in Saccharomyces cerevisiae. In: 27th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 1–4 Sept 2005, Shanghai, China
Acknowledgement
The authors thank Thomas Scheper, Technical Chemistry Institute (Institut für Technische Chemie) of Hannover University, Germany for providing the prototype of the probe, including the mechanical parts, software for controlling the electro-mechanical elements. Gratitude is also shown to the Graduate College of Bioinformatics (Graduiertenkolleg Bioinformatik) of Bielefeld University and German Research Foundation (Deutsche Forschungsgemeinschaft) for funding this project.
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Wei, N., You, J., Friehs, K. et al. In situ dark field microscopy for on-line monitoring of yeast cultures. Biotechnol Lett 29, 373–378 (2007). https://doi.org/10.1007/s10529-006-9245-x
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DOI: https://doi.org/10.1007/s10529-006-9245-x