Microscopic energy transfer spectroscopy to determine mitochondrial malfunction in human myotubes

Michael H. Gschwend; Wolfgang S. L. Strauss; H. Brinkmeier; R. Ruedel; Rudolf W. Steiner; Herbert Schneckenburger

doi:10.1117/12.260816

6 December 1996 Microscopic energy transfer spectroscopy to determine mitochondrial malfunction in human myotubes

Michael H. Gschwend, Wolfgang S. L. Strauss, H. Brinkmeier, R. Ruedel, Rudolf W. Steiner, Herbert Schneckenburger

Proceedings Volume 2926, Optical Biopsies and Microscopic Techniques; (1996) https://doi.org/10.1117/12.260816
Event: BiOS Europe '96, 1996, Vienna, Austria

Abstract

A microscopic equipment is reported for examination of cellular autofluorescence and determination of energy transfer in vitro, which is proposed to be an appropriate tool to investigate mitochondrial malfunction. The method includes fluorescence microscopy combined with time-gated (nanosecond) fluorescence emission spectroscopy and is presently used to study mitochondrial metabolism of human myotube primary cultures Enzyme complexes of the respiratory chain, located at the inner mitochondrial membrane, were inhibited by various drugs, and fluorescence of the mitochondrial coenzyme nicotinamide adenine dinucleotide (NADH) as well as of the mitochondrial marker rhodamine 123 (R123) was examined. After inhibition of enzyme complex I (NADH-coenzyme Q reductase) by rotenone or enzyme complex III (coenzyme QH₂-cytochrome c reductase) by antimycin a similar or increased NADH fluorescence was observed. In addition, energy transfer from excited states of NADH (energy donor) to R123 (energy acceptor) was deduced from a decrease of NADH fluorescence after coincubation with these inhibitors and R123. Application of microscopic energy transfer spectroscopy for diagnosis of congenital mitochondrial deficiencies is currently in preparation.

Citation Download Citation

Michael H. Gschwend, Wolfgang S. L. Strauss, H. Brinkmeier, R. Ruedel, Rudolf W. Steiner, and Herbert Schneckenburger "Microscopic energy transfer spectroscopy to determine mitochondrial malfunction in human myotubes", Proc. SPIE 2926, Optical Biopsies and Microscopic Techniques, (6 December 1996); https://doi.org/10.1117/12.260816

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