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Proceedings Paper

One- and two-component analysis of cyan fluorescent protein: FLIM-FRET microscopy
Author(s): Ye Chen; Michelle King; Ammasi Periasamy
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Paper Abstract

Remarkable advances have been made in studying the dynamic events of protein molecules in living cells and tissues using advanced light microscopy imaging techniques and green fluorescent proteins (GFPs). Identification of the interacting protein partners is critical in understanding its function and place in the biochemical pathway, thereby establishing its role in important disease processes. FLIM-FRET microscopy technique, allow the study of proteins in multiple ways including what proteins are expressed, where they are expressed- and where they move over time. It has been observed that the eCFP-eYFP FRET pair may not be that suitable to localize the association of protein molecules since the eCFP has two-components lifetime. The new Cerulean green fluorescent protein appears to have only one-component lifetime. We describe the extensive investigation of eCFP and Cerulean to study the dimerization of the transcription factor CCAAT/enhancer binding protein alpha in GHFT1-5 living cell nucleus using the time-correlated single photon counting (TCSPC) FLIM-FRET microscopy.

Paper Details

Date Published: 30 March 2005
PDF: 7 pages
Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, (30 March 2005); doi: 10.1117/12.589856
Show Author Affiliations
Ye Chen, Univ. of Virginia (United States)
Michelle King, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)

Published in SPIE Proceedings Vol. 5700:
Multiphoton Microscopy in the Biomedical Sciences V
Ammasi Periasamy; Peter T. C. So, Editor(s)

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