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Proceedings Paper

Quantitative multi-parameter analysis of single molecule dynamics by PIE FastFLIM microscopy
Author(s): Yuansheng Sun; Ulas Coskun; Phoebe S. Tsoi; Josephine C. Ferreon; Allan Chris Ferreon; Beniamino Barbieri; Shih-Chu Jeff Liao
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Paper Abstract

PIE FastFLIM microscopy allows the quantitative multi-parameter measurement of single molecule protein folding and dynamics. Using donor-acceptor FRET pair-labeled proteins, we detect changes in protein conformation and dynamics by monitoring FRET efficiency, stoichiometry and lifetime. Together with anisotropy decay information, we acquire rotational relaxation times for single molecules. By applying antibunching, FLCS and burst analysis, multi-parameters (such as copy numbers in protein complexes), diffusion coefficient and molecular brightness can be fitted for deeper understanding of the conformational dynamic behavior of single protein molecules. In this paper, we’ll focus on the multiparameters of FRET efficiency, stoichiometry and lifetime.

Paper Details

Date Published: 21 February 2017
PDF: 10 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100691X (21 February 2017); doi: 10.1117/12.2255961
Show Author Affiliations
Yuansheng Sun, ISS, Inc. (United States)
Ulas Coskun, ISS, Inc. (United States)
Phoebe S. Tsoi, Baylor College of Medicine (United States)
Josephine C. Ferreon, Baylor College of Medicine (United States)
Allan Chris Ferreon, Baylor College of Medicine (United States)
Beniamino Barbieri, ISS, Inc. (United States)
Shih-Chu Jeff Liao, ISS, Inc. (United States)


Published in SPIE Proceedings Vol. 10069:
Multiphoton Microscopy in the Biomedical Sciences XVII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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