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Issue 8, 2004
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Laser-assisted fluorescence microscopy for measuring cell membrane dynamics

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Abstract

Membranes of living cells are characterized by laser-assisted fluorescence microscopy, in particular a combination of microspectrofluorometry, total internal reflection fluorescence microscopy (TIRFM), fluorescence lifetime imaging (FLIM) and Förster resonance energy transfer (FRET) spectroscopy. The generalized polarization (GP, characterizing a spectral shift which depends on the phase of membrane lipids) as well as the effective fluorescence lifetime (τeff) of the membrane marker laurdan were revealed to be appropriate parameters for membrane stiffness and fluidity. GP decreased with temperature, but increased during cell growth and was always higher for the plasma membrane than for intracellular membranes. Microdomains of different fluorescence lifetimes τeff were observed at temperatures above 30 °C and disappeared during cell aging. Non-radiative energy transfer was used to detect laurdan selectively in close proximity to a molecular acceptor (DiI) and may present a possibility for measuring membrane dynamics in specific microenvironments.

Graphical abstract: Laser-assisted fluorescence microscopy for measuring cell membrane dynamics

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Article information


Submitted
05 Jan 2004
Accepted
17 May 2004
First published
28 May 2004

Photochem. Photobiol. Sci., 2004,3, 817-822
Article type
Paper

Laser-assisted fluorescence microscopy for measuring cell membrane dynamics

H. Schneckenburger, M. Wagner, M. Kretzschmar, W. S. L. Strauss and R. Sailer, Photochem. Photobiol. Sci., 2004, 3, 817
DOI: 10.1039/B317047K

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