scholarly journals Three-color confocal Förster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells

2015 ◽  
Vol 87 (6) ◽  
pp. 580-588 ◽  
Author(s):  
Horst Wallrabe ◽  
Yuansheng Sun ◽  
Xiaolan Fang ◽  
Ammasi Periasamy ◽  
George S. Bloom
Keyword(s):  
Energy Transfer ◽  
Quantitative Analysis ◽  
Fluorescence Resonance Energy Transfer ◽  
Protein Interactions ◽  
Actin Filaments ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Fluorescence Resonance

Imaging Protein Interactions and Gene Expression in Individual Cells by Fluorescence Resonance Energy Transfer

1999 ◽  
Vol 5 (S2) ◽  
pp. 1036-1037
Author(s):  
R.Y. Tsien ◽  
A. Miyawaki ◽  
R. Kerr ◽  
G. Baird ◽  
B.A. Griffin ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Protein Interactions ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Transgenic Animals ◽  
Dynamic Imaging ◽  
Live Cells ◽  
Fluorescence Resonance

Interactions between proteins or protein domains can be imaged by fusing them to cyan (CFP) and yellow (YFP) mutants of Green Fluorescent Protein and observing fluorescence resonance energy transfer (FRET). For example, fusions of CFP, calmodulin, a calmodulin-binding peptide, and YFP are transfectable emission-ratioing Ca2+ indicators with many uses. They are highly suitable for twophoton excitation at 770-810 nm, even at video rates. Applications not possible with previous indicators include detection of submicroscopic domains of Ca2+ by fusion of the indicators to key proteins, and dynamic imaging of Ca2+ in transgenic animals. YFPs have been improved as FRET acceptors by reducing their sensitivity to pH changes. Many other applications of GFP mutants to detect fluctuating protein-protein interactions are underway.A synthetic alternative to GFPs for protein tagging arises from the ability of membrane-permeant biarsenical dyes to seek out and light up alpha-helical Cys-Cys-X-X-Cys-Cys motifs placed in recombinant proteins in live cells. The new system is much smaller than GFP (6 residues vs. 238), can label internal domains not just N- and C-terminii, and offers novel readouts (e.g. red emission peaking > 600 nm) and better temporal control of the labeling.

Imaging protein–protein interactions in cell motility using fluorescence resonance energy transfer (FRET)

2004 ◽  
Vol 32 (3) ◽  
pp. 431-433 ◽  
Author(s):  
M. Parsons ◽  
B. Vojnovic ◽  
S. Ameer-Beg
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Conformational Changes ◽  
Protein Interactions ◽  
Intact Cell ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Fluorescence Resonance

Protein–protein interactions and signal transduction pathways have traditionally been analysed using biochemical techniques or standard microscopy. Although invaluable in the delineation of protein hierarchy, these methods do not provide information on the true spatial and temporal nature of complex formation within the intact cell. Recent advances in microscopy have allowed the development of new methods to analyse protein–protein interactions at very high resolution in both fixed and live cells. The present paper provides a brief overview of using fluorescence resonance energy transfer to analyse directly molecular interactions and conformational changes in various proteins involved in the regulation of cell adhesion and motility.

scholarly journals Spectral wide-field microscopic fluorescence resonance energy transfer imaging in live cells

2015 ◽  
Vol 20 (8) ◽  
pp. 086011 ◽  
Author(s):  
Lili Zhang ◽  
Guiqi Qin ◽  
Liuying Chai ◽  
Jiang Zhang ◽  
Fangfang Yang ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Wide Field ◽  
Fluorescence Resonance
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