A user's guide for characterizing plasma membrane subdomains in living cells by spot variation fluorescence correlation spectroscopy

2017 ◽  
pp. 1-22 ◽  
Author(s):  
S. Mailfert ◽  
Y. Hamon ◽  
N. Bertaux ◽  
H.-T. He ◽  
D. Marguet
Keyword(s):  
Plasma Membrane ◽  
Fluorescence Correlation Spectroscopy ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation

ANALYSIS OF MEMBRANE-BINDING PROTEIN MOBILITY IN LIVING CELLS USING TOTAL INTERNAL REFLECTION FLUORESCENCE CORRELATION SPECTROSCOPY

2006 ◽  
Vol 01 (03) ◽  
pp. 293-299 ◽  
Author(s):  
Y. OHSUGI ◽  
MASATAKA KINJO
Keyword(s):  
Signal Transduction ◽  
Plasma Membrane ◽  
Fluorescence Correlation Spectroscopy ◽  
Total Internal Reflection ◽  
Membrane Binding ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Fluorescence Correlation

Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) is an appropriate method for measuring diffusion constants and the number of fluorescent molecules very close to the coverglass surface. Recently, we have reported the application of TIR-FCS to cell biology, measuring membrane-binding farnesylated green fluorescent proteins (EGFP-F) in living cells. In this research, we measured the signal transduction molecule, protein kinase C (PKC), fused with EGFP in living HeLa cells by using TIR-FCS. We observed two different diffusional mobilities of PKCβII-EGFP, three-dimensional faster diffusion near the plasma membrane and slower lateral diffusion on the plasma membrane after adinosine tri phosphate (ATP) activation. These results indicate that it is possible to use TIR-FCS in the study of molecular dynamics and interactions of signal transduction proteins on the plasma membrane of the living cell.

Mobility and distribution of replication protein A in living cells using fluorescence correlation spectroscopy

2007 ◽  
Vol 82 (2) ◽  
pp. 156-162 ◽  
Author(s):  
Christophe Braet ◽  
Holger Stephan ◽  
Ian M. Dobbie ◽  
Denisio M. Togashi ◽  
Alan G. Ryder ◽  
...  
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Protein A ◽  
Replication Protein A ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Replication Protein ◽  
Fluorescence Correlation
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