Assessment of membrane potential changes using the carbocyanine dye, diS-C3-(5): synchronous excitation spectroscopy studies
Language English Country Germany Media print
Document type Journal Article
PubMed
2029874
DOI
10.1007/bf00196344
Knihovny.cz E-resources
- MeSH
- Benzothiazoles MeSH
- Potassium metabolism MeSH
- Fluorescent Dyes metabolism MeSH
- Spectrometry, Fluorescence MeSH
- Carbocyanines metabolism MeSH
- Kinetics MeSH
- Humans MeSH
- Mathematics MeSH
- Membrane Potentials * MeSH
- Leukemia, Myeloid MeSH
- Tumor Cells, Cultured MeSH
- Valinomycin pharmacology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- 3,3'-dipropyl-2,2'-thiadicarbocyanine MeSH Browser
- Benzothiazoles MeSH
- Potassium MeSH
- Fluorescent Dyes MeSH
- Carbocyanines MeSH
- Valinomycin MeSH
The fluorescence of the voltage sensitive dye, diS-C3-(5), has been analyzed by means of synchronous excitation spectroscopy. Using this rather rare fluorescence technique we have been able to distinguish between the slightly shifted spectra of diS-C3-(5) fluorescence from cells and from the supernatant. It has been found that diS-C3-(5) fluorescence in the supernatant can be selectively monitored at lambda exc = 630 nm and lambda em = 650 nm, while the cell associated fluorescence can be observed at lambda exc = 690 nm and lambda em = 710 nm. A modified theory for the diS-C3-(5) fluorescence response to the membrane potential is presented, according to which a linear relationship exists between the logarithmic increment of the dye fluorescence intensity in the supernatant, 1n I/I degrees, and the underlying change in the plasma membrane potential, delta psi p = psi p - psi p degrees. The theory has been tested on human myeloid leukemia cells (line ML-1) in which membrane potential changes were induced by valinomycin clamping in various K+ gradients. It has been demonstrated that the membrane potential change, delta psi p, can be measured on an absolute scale.
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