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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| Hauptverfasser: | , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
March 13, 2013
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| In: |
PLOS ONE
Year: 2013, Jahrgang: 8, Heft: 3 |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0058260 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1371/journal.pone.0058260 Verlag, lizenzpflichtig, Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0058260 
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| Verfasserangaben: | Richard Fairless, Andreas Beck, Mykola Kravchenko, Sarah K. Williams, Ulrich Wissenbach, Ricarda Diem, Adolfo Cavalié |
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| 520 | |a 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. | ||
| 650 | 4 | |a Cell membranes | |
| 650 | 4 | |a Fluorescence | |
| 650 | 4 | |a Fluorescence imaging | |
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| 650 | 4 | |a Membrane potential | |
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| 650 | 4 | |a Neurons | |
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