Membrane potential measurements of isolated neurons using a voltage-sensitive dye.

The ability to monitor changes in membrane potential is a useful tool for studying neuronal function, but there are only limited options available at present. Here, we have investigated the potential of a commercially available FLIPR membrane potential (FMP) dye, developed originally for high throug...

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Autores principales:	Richard Fairless, Andreas Beck, Mykola Kravchenko, Sarah K Williams, Ulrich Wissenbach, Ricarda Diem, Adolfo Cavalié
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/97e545e244264e1abaa4f02b706f66a1
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spelling	oai:doaj.org-article:97e545e244264e1abaa4f02b706f66a12021-11-18T07:53:40ZMembrane potential measurements of isolated neurons using a voltage-sensitive dye.1932-620310.1371/journal.pone.0058260https://doaj.org/article/97e545e244264e1abaa4f02b706f66a12013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23516458/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The ability to monitor changes in membrane potential is a useful tool for studying neuronal function, but there are only limited options available at present. Here, we have investigated the potential of a commercially available FLIPR membrane potential (FMP) dye, developed originally for high throughput screening using a plate reader, for imaging the membrane potential of cultured cells using an epifluorescence-based single cell imaging system. We found that the properties of the FMP dye make it highly suitable for such imaging since 1) its fluorescence displayed a high signal-to-noise ratio, 2) robust signals meant only minimal exposure times of around 5 ms were necessary, and 3) bidirectional changes in fluorescence were detectable resulting from hyper- or depolarising conditions, reaching equilibrium with a time constant of 4-8 s. Measurements were possible independently of whether membrane potential changes were induced by voltage clamping, or manipulating the ionic distribution of either Na(+) or K(+). Since FMP behaves as a charged molecule which accumulates in the cytosol, equations based on the Boltzmann distribution were developed determining that the apparent charge of FMP which represents a measure of the voltage sensitivity of the dye, is between -0.62 and -0.72. Finally, we demonstrated that FMP is suitable for use in a variety of neuronal cell types and detects membrane potential changes arising from spontaneous firing of action potentials and through stimulation with a variety of excitatory and inhibitory neurotransmitters.Richard FairlessAndreas BeckMykola KravchenkoSarah K WilliamsUlrich WissenbachRicarda DiemAdolfo CavaliéPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 3, p e58260 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Richard Fairless Andreas Beck Mykola Kravchenko Sarah K Williams Ulrich Wissenbach Ricarda Diem Adolfo Cavalié Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
description	The ability to monitor changes in membrane potential is a useful tool for studying neuronal function, but there are only limited options available at present. Here, we have investigated the potential of a commercially available FLIPR membrane potential (FMP) dye, developed originally for high throughput screening using a plate reader, for imaging the membrane potential of cultured cells using an epifluorescence-based single cell imaging system. We found that the properties of the FMP dye make it highly suitable for such imaging since 1) its fluorescence displayed a high signal-to-noise ratio, 2) robust signals meant only minimal exposure times of around 5 ms were necessary, and 3) bidirectional changes in fluorescence were detectable resulting from hyper- or depolarising conditions, reaching equilibrium with a time constant of 4-8 s. Measurements were possible independently of whether membrane potential changes were induced by voltage clamping, or manipulating the ionic distribution of either Na(+) or K(+). Since FMP behaves as a charged molecule which accumulates in the cytosol, equations based on the Boltzmann distribution were developed determining that the apparent charge of FMP which represents a measure of the voltage sensitivity of the dye, is between -0.62 and -0.72. Finally, we demonstrated that FMP is suitable for use in a variety of neuronal cell types and detects membrane potential changes arising from spontaneous firing of action potentials and through stimulation with a variety of excitatory and inhibitory neurotransmitters.
format	article
author	Richard Fairless Andreas Beck Mykola Kravchenko Sarah K Williams Ulrich Wissenbach Ricarda Diem Adolfo Cavalié
author_facet	Richard Fairless Andreas Beck Mykola Kravchenko Sarah K Williams Ulrich Wissenbach Ricarda Diem Adolfo Cavalié
author_sort	Richard Fairless
title	Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
title_short	Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
title_full	Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
title_fullStr	Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
title_full_unstemmed	Membrane potential measurements of isolated neurons using a voltage-sensitive dye.
title_sort	membrane potential measurements of isolated neurons using a voltage-sensitive dye.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/97e545e244264e1abaa4f02b706f66a1
work_keys_str_mv	AT richardfairless membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT andreasbeck membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT mykolakravchenko membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT sarahkwilliams membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT ulrichwissenbach membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT ricardadiem membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye AT adolfocavalie membranepotentialmeasurementsofisolatedneuronsusingavoltagesensitivedye
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Membrane potential measurements of isolated neurons using a voltage-sensitive dye.

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