Time-dependence of the voltage-sensitivity of second harmonic generation from styryl dyes

Andrew C. Millard; Lei Jin; Leslie M. Loew

doi:10.1117/12.646418

23 February 2006 Time-dependence of the voltage-sensitivity of second harmonic generation from styryl dyes

Andrew C. Millard, Lei Jin, Leslie M. Loew

Proceedings Volume 6089, Multiphoton Microscopy in the Biomedical Sciences VI; 60891G (2006) https://doi.org/10.1117/12.646418
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

Many styryl dyes have been developed for use in imaging electrophysiology given the fast electrochromism of their fluorescence. The molecular characteristics responsible for electrochromism also make them strong harmonophores. Second harmonic generation (SHG) from a number of these dyes has been investigated for use in a new method of imaging electrochemical activity in neurons, cardiomyocytes and other cells. Depending on the choice of dye and excitation wavelength, the voltage-sensitivity of SHG may be up to four times as large as the voltage-sensitivity of fluorescence (one- or two-photon), as is well characterised by "slow" voltage-switching experiments. While some fluorescent dyes respond to changes in trans-membrane potential by slow mechanisms, such as a change in partitioning between the cell membrane and the extracellular medium, styryl dyes were developed because they respond rapidly, using a fast electrochromic shift of their fluorescence excitation and emission spectra. For use in imaging mammalian nerves, for instance, a response time of the order of a millisecond is necessary. Here we report on our characterisation of the time dependence of the voltage-sensitivity of SHG from styryl dyes using "fast" voltage-switching experiments, as compared with simultaneous two-photon fluorescence imaging for a number of different dyes.

Citation Download Citation

Andrew C. Millard, Lei Jin, and Leslie M. Loew "Time-dependence of the voltage-sensitivity of second harmonic generation from styryl dyes", Proc. SPIE 6089, Multiphoton Microscopy in the Biomedical Sciences VI, 60891G (23 February 2006); https://doi.org/10.1117/12.646418

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