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Single‐ and two‐photon spectral imaging of intrinsic fluorescence of transformed human hepatocytes
Author(s) -
Rajwa Bartek,
Bernas Tytus,
Acker Helmut,
Dobrucki Jurek,
Robinson J.P.
Publication year - 2007
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20497
Subject(s) - autofluorescence , membrane , biophysics , chemistry , two photon excitation microscopy , fluorescence , cytoplasm , in vitro , cell membrane , biochemistry , biology , optics , physics
Autofluorescence (AF) originating from the cytoplasmic region of mammalian cells has been thoroughly investigated; however, AF from plasma membranes of viable intact cells is less well known, and has been mentioned only in a few older publications. Herein, we report results describing single‐ and two‐photon spectral properties of a strong yellowish‐green AF confined to the plasma‐membrane region of transformed human hepatocytes (HepG2) grown in vitro as small three‐dimensional aggregates or as monolayers. The excitation–emission characteristics of the membrane AF indicate that it may originate from a flavin derivative. Furthermore, the AF was closely associated with the plasma membranes of HepG2 cells, and its presence and intensity were dependent on cell metabolic state, membrane integrity and presence of reducing agents. This AF could be detected both in live intact cells and in formaldehyde‐fixed cells. Microsc. Res. Tech., 2007. © 2007 Wiley‐Liss, Inc.