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Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid-protein interactions and the traditional lipid 'raft' theory.

Original publication

DOI

10.1042/bse0570069

Type

Journal article

Journal

Essays Biochem

Publication Date

2015

Volume

57

Pages

69 - 80

Keywords

Diffusion, Fluorescent Dyes, Lipid Bilayers, Membrane Microdomains, Membrane Proteins, Microscopy, Confocal, Phosphatidylcholines, Phosphatidylethanolamines, Spectrometry, Fluorescence, Sphingomyelins