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Journal of Biomedical Optics

Time-resolved in situ measurement of mitochondrial malfunction by energy transfer spectroscopy
Author(s): Herbert Schneckenburger; Reinhard Sailer; Wolfgang S. L. Strauss; Marco Lyttek; Karl Stock; Peter Zipfl
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Paper Abstract

To establish optical in situ detection of mitochondrial malfunction, nonradiative 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. A screening system was developed in which these signals are detected simultaneously using a time-gated (nanosecond) 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 results obtained from cultivated endothelial cells are reported. The ratio of fluorescence intensities excited in the near ultraviolet and blue–green spectral ranges increased by a factor 1.5 or 1.35 after inhibition of the mitochondrial respiratory chain by rotenone at cytotoxic or noncytotoxic concentrations, respectively. Concomitantly the amount of mitochondrial NADH increased. Excellent linearity between the number of cells incubated with R123 and fluorescence intensity was found in suspension.

Paper Details

PDF: 5 pages
J. Biomed. Opt. 5(4) doi: 10.1117/12.351027
Published in: Journal of Biomedical Optics Volume 5, Issue 4, October 2000
Show Author Affiliations
Herbert Schneckenburger, Fachhochschule Aalen (Germany)
Reinhard Sailer, Univ. Ulm (Germany)
Wolfgang S. L. Strauss, Univ. Ulm (Germany)
Marco Lyttek, Inst. fur Angewandte Forschung (Germany)
Karl Stock, Univ. Ulm (Germany)
Peter Zipfl, Fachhochschule Aalen (Germany)


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