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Proceedings Paper

Studies of the metabolism of cell cultures by microspectrofluoroscopy
Author(s): Wolfgang Hoehne; Werner Schramm; V. Moritzen; U. Burgmann; Heinz Detlef Kronfeldt
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Paper Abstract

The monitoring of the state of cellular energy metabolism and respiratory activity is a necessary procedure in cell biology and pharmacology. One method is the observation of the redox state by NADH and FAD autofluorescence measurements. Using this technique, investigations on endothelial cell cultures were done to study their behavior under pharmacologic influences. One application was the investigation of cytotoxicity of cyanides, blocking the mitochondrial respiratory chain. Further we studied the activation of energy metabolism as a step of the cellular reaction on extracellular impacts. The measurements have been performed with a fluorescence microscope Zei(beta) Axioplan, extended by a PMT and a CCD camera. During examination, the cell cultures were kept under nearly physiological conditions using a specialized perfusion chamber. The measurements took place on cellular monolayers. Different excitation geometries have been studied to overcome the difficulties, which arose from the very weak absorption of the cell monolayer, resulting in a low quantum yield and SNR. In classical cytotoxicity studies, only the statistical long-time effects (e.g. IC50) of cell damages are recorded. By redox microspectrofluorometry it is possible to observe the process of damage in its progress, shown by the presented results. In the second, more complex model, we studied the reaction of cells on ligands like PIA (Phenylisopropyladenosin). In this case, the intracellular reaction is connected with an increased production of cAMP. Again, this requires an increased production of ATP, which leads to an activation of the cellular energy metabolism. The spectroscopic results are interpreted by a first model.

Paper Details

Date Published: 15 January 1996
PDF: 5 pages
Proc. SPIE 2628, Optical and Imaging Techniques for Biomonitoring, (15 January 1996); doi: 10.1117/12.229980
Show Author Affiliations
Wolfgang Hoehne, Institute of Molecular Pharmacology (Germany)
Werner Schramm, Institute of Molecular Pharmacology (Germany)
V. Moritzen, Technical Univ. Berlin (Germany)
U. Burgmann, Technical Univ. Berlin (Germany)
Heinz Detlef Kronfeldt, Technical Univ. Berlin (Germany)


Published in SPIE Proceedings Vol. 2628:
Optical and Imaging Techniques for Biomonitoring
Hans-Jochen Foth; Renato Marchesini; Halina Podbielska; Michel Robert-Nicoud; Herbert Schneckenburger, Editor(s)

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