Microspectrofluorometry and polarization microscopy of membrane dynamics in living cells

Michael Wagner; Petra Weber; Herbert Schneckenburger

doi:10.1117/12.662023

14 April 2006 Microspectrofluorometry and polarization microscopy of membrane dynamics in living cells

Michael Wagner, Petra Weber, Herbert Schneckenburger

Proceedings Volume 6191, Biophotonics and New Therapy Frontiers; 619109 (2006) https://doi.org/10.1117/12.662023
Event: SPIE Photonics Europe, 2006, Strasbourg, France

Abstract

Membrane dynamics of human glioblastoma cells were investigated using the intercalating fluorescence marker 6-dodecanoyl-2-dimethylamino naphthalene (laurdan). In particular its generalized polarization (GP), which describes a spectral shift depending on the phase of membrane lipids, was used as a measure of membrane stiffness, whereas its fluorescence lifetime τ and its rotational diffusion time t_r were used to characterize membrane fluidity. Upon excitation with linearly polarized pulsed laser light the parallel and perpendicular components of fluorescence from the sample were measured simultaneously using an imaging device with polarization sensitivity. So far, membrane dynamics depended on temperature and cell age as well as the on intracellular amount of cholesterol. In addition, the plasma membrane (assessed by illumination with an evanescent electromagnetic wave) appeared to be stiffer than intracellular membranes (assessed by epiillumination of the cells).

Citation Download Citation

Michael Wagner, Petra Weber, and Herbert Schneckenburger "Microspectrofluorometry and polarization microscopy of membrane dynamics in living cells", Proc. SPIE 6191, Biophotonics and New Therapy Frontiers, 619109 (14 April 2006); https://doi.org/10.1117/12.662023

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