One- and two-component analysis of cyan fluorescent protein: FLIM-FRET microscopy

Ye Chen; Michelle King; Ammasi Periasamy

doi:10.1117/12.589856

30 March 2005 One- and two-component analysis of cyan fluorescent protein: FLIM-FRET microscopy

Ye Chen, Michelle King, Ammasi Periasamy

Proceedings Volume 5700, Multiphoton Microscopy in the Biomedical Sciences V; (2005) https://doi.org/10.1117/12.589856
Event: SPIE BiOS, 2005, San Jose, CA, United States

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.

Citation Download Citation

Ye Chen, Michelle King, and Ammasi Periasamy "One- and two-component analysis of cyan fluorescent protein: FLIM-FRET microscopy", Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, (30 March 2005); https://doi.org/10.1117/12.589856

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KEYWORDS

Molecules

Proteins

Luminescence

Fluorescence resonance energy transfer

Microscopy

Green fluorescent protein

Microscopes

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