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Proceedings Paper

Combined fluorescence and ultrastructural mapping of living cells
Author(s): Elli Kohen; Joseph G. Hirschberg; Cahide Kohen; Jeffrey Prince; Szymon Suckewer; Rene C. Santus; Patrice Morliere; Louis Dubertret
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Paper Abstract

The topographic analysis of fluorescence distribution has been carried out pixel-by-pixel by one dimensional, two-dimensional microspectrofluorometry and three-dimensional confocal fluorescence microscopy. Fluorescence emission spectra of NAD(P)H and benzo(a)pyrene (or metabolites) were recorded at different excitation wavelengths. Cell bioenergetics are monitored in normal and malignant cells as well as cells with genetic defects by coenzyme responses to microinjections of substrates and modifiers from key metabolic pathways in presence and absence of inhibitors and drugs active on mitochondrial structure and function. Cooperative interactions between organelles involved in detoxification mechanisms are observed in cells treated with fluorescent cytotoxic agents. Such interactions can be directly mapped by the fluorescence of cytotoxic agents, their reaction products or vital probes such as NBD ceramide for the Golgi apparatus. To identify the organelles involved parallel electron microscopic studies are carried out in cells first treated with the cytotoxic agent and then incubated with an electron opaque material. A recently developed combined X-ray laser microscope (COXRALM) holds the promise of carrying out combined phase-fluorescence-and X-ray microscopic observations of fluorescence and ultrastructural correlations in live cell probing. As further versatility is gained in such methods it may become possible to obtain a very detailed structure and function mapping of living cells within the context of cytomatrix analysis, metabolic compartmentation and organelle interactions.

Paper Details

Date Published: 1 May 1990
PDF: 17 pages
Proc. SPIE 1204, Time-Resolved Laser Spectroscopy in Biochemistry II, (1 May 1990); doi: 10.1117/12.17719
Show Author Affiliations
Elli Kohen, Univ. of Miami (United States)
Joseph G. Hirschberg, Univ. of Miami (United States)
Cahide Kohen, Univ. of Miami (United States)
Jeffrey Prince, Univ. of Miami (United States)
Szymon Suckewer, Princeton Univ. (United States)
Rene C. Santus, Museum National d'Histoire Naturelle (France)
Patrice Morliere, Hopital Henri Mondor/INSERM (France)
Louis Dubertret, Hopital Henri Mondor/INSERM (France)

Published in SPIE Proceedings Vol. 1204:
Time-Resolved Laser Spectroscopy in Biochemistry II
Joseph R. Lakowicz, Editor(s)

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