Time-resolved in-situ measurement of mitochondrial malfunction by energy transfer spectroscopy

Herbert Schneckenburger; Michael H. Gschwend; Wolfgang S. L. Strauss; Reinhard Sailer; Lars Schoch; Alexander Schuh; Karl Stock; Rudolf W. Steiner; Peter Zipfl

doi:10.1117/12.351027

2 July 1999 Time-resolved in-situ measurement of mitochondrial malfunction by energy transfer spectroscopy

Herbert Schneckenburger, Michael H. Gschwend, Wolfgang S. L. Strauss, Reinhard Sailer, Lars Schoch, Alexander Schuh, Karl Stock, Rudolf W. Steiner, Peter Zipfl

Author Affiliations +

Proceedings Volume 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation; (1999) https://doi.org/10.1117/12.351027
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

To establish optical in situ detection of mitochondrial malfunction, non-radiative energy transfer from the coenzyme NADH to the mitochondrial marker rhodamine 123 (R123) was examined. Dual excitation of R123 via energy transfer from excited NADH molecules as well as by direct absorption of light results in two fluorescence signals whose ratio is a measure of mitochondrial NADH. An experimental setup was developed, where these signals are detected simultaneously using a time-gated technique for energy transfer measurements and a frequency selective technique for direct excitation and fluorescence monitoring of R123. Optical and electronic components of the apparatus are described, and preliminary result of cultivated endothelial cells are reported. Results are compared with those obtained from a previously established microscopic system and discussed in view of potential applications.

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Herbert Schneckenburger, Michael H. Gschwend, Wolfgang S. L. Strauss, Reinhard Sailer, Lars Schoch, Alexander Schuh, Karl Stock, Rudolf W. Steiner, and Peter Zipfl "Time-resolved in-situ measurement of mitochondrial malfunction by energy transfer spectroscopy", Proc. SPIE 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation, (2 July 1999); https://doi.org/10.1117/12.351027

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