TECHNICAL PAPERS

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

[+] Author Affiliations
Herbert Schneckenburger

Institut fu¨r Lasertechnologien in der Medizin und Messtechnik an der Universita¨t Ulm, Ulm, Germany,

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, Aalen, Germany,

Michael H. Gschwend

Universita¨t Ulm, Abteilung fu¨r Allgemeine Physiologie, Ulm, Germany

Reinhard Sailer

Institut fu¨r Lasertechnologien in der Medizin und Messtechnik an der Universita¨t Ulm, Ulm, Germany

Wolfgang S. L. Strauss

Institut fu¨r Lasertechnologien in der Medizin und Messtechnik an der Universita¨t Ulm, Ulm, Germany

Marco Lyttek

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, Aalen, Germany

Karl Stock

Institut fu¨r Lasertechnologien in der Medizin und Messtechnik an der Universita¨t Ulm, Ulm, Germany,

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, Aalen, Germany

Peter Zipfl

Institut fu¨r Angewandte Forschung, Fachhochschule Aalen, Aalen, Germany

J. Biomed. Opt. 5(4), 362-366 (Oct 01, 2000). doi:10.1117/1.1289358
History: Received Nov. 1, 1999; Revised Apr. 11, 2000; Revised June 26, 2000; Accepted June 26, 2000
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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. © 2000 Society of Photo-Optical Instrumentation Engineers.

© 2000 Society of Photo-Optical Instrumentation Engineers

Citation

Herbert Schneckenburger ; Michael H. Gschwend ; Reinhard Sailer ; Wolfgang S. L. Strauss ; Marco Lyttek, et al.
"Time-resolved in situ measurement of mitochondrial malfunction by energy transfer spectroscopy", J. Biomed. Opt. 5(4), 362-366 (Oct 01, 2000). ; http://dx.doi.org/10.1117/1.1289358


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