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Direct visualization of the dynamics of membrane‐anchor proteins in living cells
Author(s) -
WANG C.,
FU G.,
WANG J.,
WANG G.,
CHENG Y.,
XU Z.Z.
Publication year - 2008
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2007.01865.x
Subject(s) - cytoplasm , total internal reflection fluorescence microscope , biophysics , membrane , fluorescence microscope , fluorescence , cell membrane , membrane protein , chemistry , microbiology and biotechnology , tracking (education) , fluorescence lifetime imaging microscopy , biology , biochemistry , physics , optics , psychology , pedagogy
Summary Dynamic properties of proteins have crucial roles in understanding protein function and molecular mechanism within cells. In this paper, we combined total internal reflection fluorescence microscopy with oblique illumination fluorescence microscopy to observe directly the movement and localization of membrane‐anchored green fluorescence proteins in living cells. Total internal reflect illumination allowed the observation of proteins in the cell membrane of living cells since the penetrate depth could be adjusted to about 80 nm, and oblique illumination allowed the observation of proteins both in the cytoplasm and apical membrane, which made this combination a promising tool to investigate the dynamics of proteins through the whole cell. Not only individual protein molecule tracks have been analyzed quantitatively but also cumulative probability distribution function analysis of ensemble trajectories has been done to reveal the mobility of proteins. Finally, single particle tracking has acted as a compensation for single molecule tracking. All the results exhibited green fluorescence protein dynamics within cytoplasm, on the membrane and from cytoplasm to plasma membrane.