Share Email Print
cover

Proceedings Paper

Influence of the refractive index on EGFP fluorescence lifetimes in mixtures of water and glycerol
Author(s): Klaus Suhling; Daniel M. Davis; Zdenek Petrasek; Jan Siegel; David Phillips
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper Abstract

As a precursor to applying fluorescence lifetime imaging (FLIM) to studies of intercellular communication in molecular immunology, we have investigated the fluorescence lifetime of enhanced green fluorescent protein (EGFP) in mixtures of water and glycerol using time-correlated single photon counting (TCSPC). We find that the EGFP lifetime decreases with increasing glycerol content. This is accounted for quantitatively by the refractive index dependence of the fluorescence lifetime as predicted by the Strickler Berg formula which relates the fluorescence lifetime to the absorption spectrum. The solvent viscosity has no influence on the fluorescence lifetime. We also discuss the refractive index dependence of the GFP fluorescence lifetime in more complex systems. The findings are particularly relevant for the interpretation of FLIM of GFP expressed in environments such as bacteria and cells.

Paper Details

Date Published: 2 July 2001
PDF: 10 pages
Proc. SPIE 4259, Biomarkers and Biological Spectral Imaging, (2 July 2001); doi: 10.1117/12.432486
Show Author Affiliations
Klaus Suhling, Imperial College of Science, Technology and Medicine (United Kingdom)
Daniel M. Davis, Imperial College of Science, Technology and Medicine (United Kingdom)
Zdenek Petrasek, Imperial College of Science, Technology and Medicine (United Kingdom)
Jan Siegel, Imperial College of Science, Technology and Medicine (United Kingdom)
David Phillips, Imperial College of Science, Technology and Medicine (United Kingdom)


Published in SPIE Proceedings Vol. 4259:
Biomarkers and Biological Spectral Imaging
Gregory H. Bearman; Darryl J. Bornhop; Richard M. Levenson; Darryl J. Bornhop, Editor(s)

© SPIE. Terms of Use
Back to Top