High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge

Sensitivity spectra of Stark-shift voltage sensitive dyes, such as ANNINE-6, suggest the use of the extreme red edges of the excitation spectrum to achieve large fractional fluorescence changes with membrane voltage. This was tested in cultured HEK293 cells. Cells were illuminated with light at the...

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Main Authors: Kuhn, Bernd, Fromherz, Peter, Denk, Winfried
Formato: Artigo
Idioma:Inglês
Publicado em: Biophysical Society 2004
Assuntos:
Spectroscopy, Imaging, Other Techniques
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC1304385/
https://ncbi.nlm.nih.gov/pubmed/15240496
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1529/biophysj.104.040477
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spelling pubmed-13043852006-01-17 High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge Kuhn, Bernd Fromherz, Peter Denk, Winfried Biophys J Spectroscopy, Imaging, Other Techniques Sensitivity spectra of Stark-shift voltage sensitive dyes, such as ANNINE-6, suggest the use of the extreme red edges of the excitation spectrum to achieve large fractional fluorescence changes with membrane voltage. This was tested in cultured HEK293 cells. Cells were illuminated with light at the very red edge of the dye's excitation spectrum, where the absorption cross section is as much as 100 times smaller than at its peak. The small-signal fractional fluorescence changes were −0.17%/mV, −0.28%/mV, and −0.35%/mV for one-photon excitation at 458 nm, 488 nm, and 514 nm, respectively, and −0.29%/mV, −0.43%/mV, and −0.52%/mV for two-photon excitation at 960 nm, 1000 nm, and 1040 nm, respectively. For large voltage swings the fluorescence changes became nonlinear, reaching 50% and −28% for 100 mV hyper- and depolarization, respectively, at 514 nm and 70% and −40% at 1040 nm. Such fractional sensitivities are ∼5 times larger than what is commonly found with other voltage-sensing dyes and approach the theoretical limit given by the spectral Boltzmann tail. Biophysical Society 2004-07 /pmc/articles/PMC1304385/ /pubmed/15240496 http://dx.doi.org/10.1529/biophysj.104.040477 Text en Copyright © 2004, Biophysical Society
institution US NLM
collection PubMed Central
language Inglês
format Artigo
topic Spectroscopy, Imaging, Other Techniques
spellingShingle Spectroscopy, Imaging, Other Techniques
Kuhn, Bernd
Fromherz, Peter
Denk, Winfried
High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
description Sensitivity spectra of Stark-shift voltage sensitive dyes, such as ANNINE-6, suggest the use of the extreme red edges of the excitation spectrum to achieve large fractional fluorescence changes with membrane voltage. This was tested in cultured HEK293 cells. Cells were illuminated with light at the very red edge of the dye's excitation spectrum, where the absorption cross section is as much as 100 times smaller than at its peak. The small-signal fractional fluorescence changes were −0.17%/mV, −0.28%/mV, and −0.35%/mV for one-photon excitation at 458 nm, 488 nm, and 514 nm, respectively, and −0.29%/mV, −0.43%/mV, and −0.52%/mV for two-photon excitation at 960 nm, 1000 nm, and 1040 nm, respectively. For large voltage swings the fluorescence changes became nonlinear, reaching 50% and −28% for 100 mV hyper- and depolarization, respectively, at 514 nm and 70% and −40% at 1040 nm. Such fractional sensitivities are ∼5 times larger than what is commonly found with other voltage-sensing dyes and approach the theoretical limit given by the spectral Boltzmann tail.
author Kuhn, Bernd
Fromherz, Peter
Denk, Winfried
author_facet Kuhn, Bernd
Fromherz, Peter
Denk, Winfried
author_sort Kuhn, Bernd
title High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
title_short High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
title_full High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
title_fullStr High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
title_full_unstemmed High Sensitivity of Stark-Shift Voltage-Sensing Dyes by One- or Two-Photon Excitation Near the Red Spectral Edge
title_sort high sensitivity of stark-shift voltage-sensing dyes by one- or two-photon excitation near the red spectral edge
publisher Biophysical Society
publishDate 2004
url https://ncbi.nlm.nih.gov/pmc/articles/PMC1304385/
https://ncbi.nlm.nih.gov/pubmed/15240496
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1529/biophysj.104.040477
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