Time-resolved spectroscopy of endogenous NAD(P)H in Gluconobacter oxydans

J. Horilova; K. Kromkova; M. Bucko; A. Illesova; A. Vikartovska; V. Stefuca; A. Mateasik; D. Chorvat Jr.; A. Chorvatova

doi:10.1117/12.2001368

22 February 2013 Time-resolved spectroscopy of endogenous NAD(P)H in Gluconobacter oxydans

J. Horilova, K. Kromkova, M. Bucko, A. Illesova, A. Vikartovska, V. Stefuca, A. Mateasik, D. Chorvat Jr., A. Chorvatova

Author Affiliations +

Proceedings Volume 8588, Multiphoton Microscopy in the Biomedical Sciences XIII; 85880V (2013) https://doi.org/10.1117/12.2001368
Event: SPIE BiOS, 2013, San Francisco, California, United States

Abstract

The genus Gluconobacter is frequently used for biotechnological and/or nanotechnological applications. We studied endogenous fluorescence of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), indicator of the oxidative metabolic state in mammalian cells, in Gluconobacter oxydans (G. oxydans). Time-resolved measurements (excitation by 375nm pulsed diode laser) were employed to record the bacterial fluorescence intensity, as well as its modifications by metabolic modulation. Results were gathered on fresh bacteria, on de-frozen ones, as well as on bacteria encapsulated in alginate beads. NAD(P)H fluorescence increased linearly with the concentration of bacteria. Freezing, which has little effect on the viability of bacteria or the concentration-dependent fluorescence rise, affected the temperature-dependence of NAD(P)H fluorescence. Sodium cyanide (10 mM) provoked significant rise in the NAD(P)H fluorescence, while dinitrophenol (200 μM) induced its decrease, confirming the bacterial NAD(P)H fluorescence sensitivity to modulators of electron transport chain. Gathered results demonstrate that endogenous NAD(P)H fluorescence can be successfully recorded in the bacterial strain G. oxydans using time-resolved measurements.

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J. Horilova, K. Kromkova, M. Bucko, A. Illesova, A. Vikartovska, V. Stefuca, A. Mateasik, D. Chorvat Jr., and A. Chorvatova "Time-resolved spectroscopy of endogenous NAD(P)H in Gluconobacter oxydans", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85880V (22 February 2013); https://doi.org/10.1117/12.2001368

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