Near-membrane protein dynamics revealed by evanescent field microscopy

Vassilios J. Bezzerides; David E. Clapham

doi:10.1117/12.548399

25 May 2004 Near-membrane protein dynamics revealed by evanescent field microscopy

Vassilios J. Bezzerides, David E. Clapham

Proceedings Volume 5467, Fluctuations and Noise in Biological, Biophysical, and Biomedical Systems II; (2004) https://doi.org/10.1117/12.548399
Event: Second International Symposium on Fluctuations and Noise, 2004, Maspalomas, Gran Canaria Island, Spain

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.

Citation Download Citation

Vassilios J. Bezzerides and David E. Clapham "Near-membrane protein dynamics revealed by evanescent field microscopy", Proc. SPIE 5467, Fluctuations and Noise in Biological, Biophysical, and Biomedical Systems II, (25 May 2004); https://doi.org/10.1117/12.548399

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