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Proceedings Paper

Enhanced energy transfer in respiratory-deficient endothelial cells probed by microscopic fluorescence excitation spectroscopy
Author(s): Herbert Schneckenburger; Michael H. Gschwend; Manfred Bauer; Wolfgang S. L. Strauss; Rudolf W. Steiner
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Paper Abstract

Mitochondrial malfunction may be concomitant with changes of the redox states of the coenzymes nicotinamide adenine dinucleotide (NAD+/NADH), as well as flavin.mononucleotide or dinucleotide. The intrinsic fluorescence of these coenzymes was therefore proposed to be a measure of malfunction. Since mitochondrial fluorescence is strongly superposed by autofluorescence from various cytoplasmatic fluorophores, cultivated endothelial cells were incubated with the mitochondrial marker rhodamine 123 (R123), and after excitation of flavin molecules, energy transfer to R123 was investigated. Due to spectral overlap of flavin and R123 fluorescence, energy transfer flavin yields R123 could not be detected from their emission spectra. Therefore, the method of microscopic fluorescence excitation spectroscopy was established. When detecting R123 fluorescence, excitation maxima at 370 - 390 nm and 420-460 nm were assigned to flavins, whereas a pronounced excitation band at 465 - 490 nm was attributed to R123. Therefore, excitation at 475 nm reflected the intracellular concentration of R123, whereas excitation at 385 nm reflected flavin excitation with a subsequent energy transfer to R123 molecules. An enhanced energy transfer after inhibition of specific enzyme complexes of the respiratory chain is discussed in the present article.

Paper Details

Date Published: 6 December 1996
PDF: 9 pages
Proc. SPIE 2926, Optical Biopsies and Microscopic Techniques, (6 December 1996); doi: 10.1117/12.260811
Show Author Affiliations
Herbert Schneckenburger, Univ. Ulm and Fachhochschule Aalen (Germany)
Michael H. Gschwend, Univ. Ulm (Germany)
Manfred Bauer, Fachhochschule Aalen (Germany)
Wolfgang S. L. Strauss, Univ. Ulm (Germany)
Rudolf W. Steiner, Univ. Ulm (Germany)

Published in SPIE Proceedings Vol. 2926:
Optical Biopsies and Microscopic Techniques
Irving J. Bigio; Warren S. Grundfest; Herbert Schneckenburger; Katarina Svanberg; Pierre M. Viallet, Editor(s)

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