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Proceedings Paper

Near-membrane protein dynamics revealed by evanescent field microscopy
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Paper Abstract

Evanescent Field (EF) microscopy is used to investigate the spatial and temporal dynamics of proteins in living cells. A genetically engineered ion channel fused to a fluorescent tag is expressed in cells and imaged with an objective-based EF microscope. Images are obtained from a CCD and analyzed to determine fluorescence and velocity of individual protein containing vesicles. An inverse correlation between fluorescent intensity and average motility provides a method for determination of membrane localization. Stimulation and subsequent decrease in ion channel activity is correlated with loss of protein from membrane as shown by EF microscopy and patch-clamp electrophysiology.

Paper Details

Date Published: 25 May 2004
PDF: 12 pages
Proc. SPIE 5467, Fluctuations and Noise in Biological, Biophysical, and Biomedical Systems II, (25 May 2004); doi: 10.1117/12.548399
Show Author Affiliations
Vassilios J. Bezzerides, Harvard Medical School (United States)
Children's Hospital Boston (United States)
David E. Clapham, Harvard Medical School (United States)
Children's Hospital Boston (United States)


Published in SPIE Proceedings Vol. 5467:
Fluctuations and Noise in Biological, Biophysical, and Biomedical Systems II
Derek Abbott; Sergey M. Bezrukov; Andras Der; Angel Sanchez, Editor(s)

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