In vivo measurement of F420 fluorescence in cultures of Methanobacterium thermoautotrophicum |
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| Institution: | 1. Gesellschaft für Strahlen- und Umweltforschung München, Institut für Angewandte Optik, D-8042 Neuherberg, F.R.G.;2. Kernforschungsanlage Jülich, Institut für Biotechnologie, D-5170 Jülich, F.R.G.;1. Department of Urology, Faculty of Medicine, University of Freiburg Medical Centre, Freiburg, Germany;2. Becker & Hickl GmbH, Berlin, Germany;1. Evergreen Health Hospitalist Services, Kirkland, Washington;2. Urology Section, Department of Surgery, Veterans Affairs Medical Centers, Durham, North Carolina;3. Department of Biostatistics and Bioinformatics, Duke University School of Medicine, Durham, North Carolina;4. Department of Urology, University of California-Los Angeles Medical Center, Los Angeles, California;5. Department of Surgery, Division of Urology and Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California;6. Department of Urology, University of California-San Diego Medical Center, San Diego, California;7. Department of Urology, University of California-San Francisco Helen Diller Family Comprehensive Cancer Center, San Francisco, California;8. Urology Section, Division of Surgery, Veterans Affairs Medical Center and Division of Urologic Surgery, Department of Surgery, Medical College of Georgia, Augusta, Georgia;9. Department of Urology, Oregon Health and Science University, Portland, Oregon;1. Synthetic Single Crystal Research Center, CAS Key Laboratory of Transparent and Opto-functional Inorganic Materials, Shanghai Institute of Ceramics, Chinese Academy of Sciences, No. 588 Heshuo Road, Shanghai 201899, China;2. State Key Laboratory on High Power Semiconductor Lasers, Changchun University of Science and Technology, No. 7989 Weixing Road, Changchun 130022, Jilin, China;3. School of Physics Science and Engineering, Institute for Advanced Study, Tongji University, Shanghai 200092, China;4. University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China;5. State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics Chinese Academy of Sciences, Shanghai, 20050, China |
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| Abstract: | Growth of Methanobacterium thermoautotrophicum could be followed by measuring the intensity of fluorescence directly in the culture vessel, avoiding conventional time-consuming extraction procedures of fluorescent coenzymes. The influence of light scattering by the bacteria was investigated. It could be shown, that light scattering had only little effect on the measurement of coenzyme F420 fluorescence. However, culture fluorescence did not correlate to methanogenic activity, due to superposition of bacterial fluorescence by fluorescence from cell-free coenzyme which accumulates in the culture medium. By use of time-resolved laser spectroscopy, different fluorescence lifetimes were obtained for intracellular (1.0 ns) and extracellular (2.5 ns) components, respectively. A combination of this technique with photobleaching measurements for direct determination of F420 content of bacteria in a culture is proposed. |
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