Influence of the refractive index on EGFP fluorescence lifetimes in mixtures of water and glycerol

Klaus Suhling; Daniel M. Davis; Zdenek Petrasek; Jan Siegel; David Phillips

doi:10.1117/12.432486

2 July 2001 Influence of the refractive index on EGFP fluorescence lifetimes in mixtures of water and glycerol

Klaus Suhling, Daniel M. Davis, Zdenek Petrasek, Jan Siegel, David Phillips

Proceedings Volume 4259, Biomarkers and Biological Spectral Imaging; (2001) https://doi.org/10.1117/12.432486
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

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.

Citation Download Citation

Klaus Suhling, Daniel M. Davis, Zdenek Petrasek, Jan Siegel, and David Phillips "Influence of the refractive index on EGFP fluorescence lifetimes in mixtures of water and glycerol", Proc. SPIE 4259, Biomarkers and Biological Spectral Imaging, (2 July 2001); https://doi.org/10.1117/12.432486

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