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7668895 
Journal Article 
Assessment of membrane potential changes using the carbocyanine dye, diS-C3-(5): synchronous excitation spectroscopy studies 
Plásek, J; Hrouda, V 
1991 
Yes 
European Biophysics Journal
ISSN: 0175-7571
EISSN: 1432-1017 
Springer-Verlag 
19 
4 
183-188 
English 
2029874 
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. 
diS-C3(5); Fluorescence; Membrane potential; Synchronous excitation spectroscopy