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Biophys J. 2004 Feb;86(2):1169-76.

Sensitivity of second harmonic generation from styryl dyes to transmembrane potential.

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1
Department of Physiology and Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, Connecticut 06030-1507, USA.

Abstract

In this article we present results from the simultaneous nonlinear (second harmonic generation and two-photon excitation fluorescence) imaging and voltage clamping of living cells. Specifically, we determine the sensitivity to transmembrane potential of second harmonic generation by ANEP-chromophore styryl dyes as a function of excitation wavelength and dye structure. We have measured second harmonic sensitivities of up to 43% per 100 mV, more than a factor of four better than the nominal voltage sensitivity of the dyes under "one-photon" fluorescence. We find a dependence of voltage sensitivity on excitation wavelength that is consistent with a two-photon resonance, and there is a significant dependence of voltage sensitivity on the structure of the nonchromophore portion of the dyes.

PMID:
14747351
PMCID:
PMC1303909
DOI:
10.1016/S0006-3495(04)74191-0
[Indexed for MEDLINE]
Free PMC Article
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