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Proceedings Paper

Comparison of FRET microscopy imaging techniques for studying protein-protein interactions in living cells using FRET standards
Author(s): Yuansheng Sun; Soo-Ah Seo; Sydney Provence; Ammasi Periasamy
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Paper Abstract

Förster resonance energy transfer (FRET) microscopy is a powerful tool to localize protein-protein interactions in living specimens. Various FRET microscopy imaging techniques have been established and are generally categorized into intensity-based and lifetime-based methods. Based on the detection of the acceptor sensitized emission, we have developed the FRET imaging methodologies that can be applied in combination of wide-field, conventional confocal or spectral microscopy. All these FRET microscopy methods have the capability to interpret the change in proximity between the donor and the acceptor through measuring the apparent energy transfer efficiency (E). However, to answer what subtle change of E can be detected and to link correctly FRET data to biological information, the imaging techniques have to be well calibrated. In this regard, we compare various FRET microscopy methods and assess their utilities using several genetic ("FRET standard") constructs where Cerulean and Venus fluorescent proteins are tethered by different amino acid linkers.

Paper Details

Date Published: 26 February 2010
PDF: 10 pages
Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75690Z (26 February 2010); doi: 10.1117/12.842186
Show Author Affiliations
Yuansheng Sun, Univ. of Virginia (United States)
Soo-Ah Seo, Univ. of Virginia (United States)
Sydney Provence, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)

Published in SPIE Proceedings Vol. 7569:
Multiphoton Microscopy in the Biomedical Sciences X
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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