The fluorescent dye 3,3'-dipropylthiadicarbocyanine, diS-C(3)(3), is a suitable probe to monitor real changes of plasma membrane potential in yeast cells which are too small for direct membrane potential measurements with microelectrodes. A method presented in this paper makes it possible to convert changes of equilibrium diS-C(3)(3) fluorescence spectra, measured in yeast cell suspensions under certain defined conditions, into underlying membrane potential differences, scaled in the units of millivolts. Spectral analysis of synchronously scanned diS-C(3)(3) fluorescence allows to assess the amount of dye accumulated in cells without otherwise necessary sample taking and following separation of cells from the medium. Moreover, membrane potential changes can be quantified without demanding calibration protocols. The applicability of this approach was demonstrated on the depolarization of Rhodotorula glutinis yeast cells upon acidification of cell suspensions and/or by increasing extracellular K(+) concentration.

Charles University Prague Prague Czech Republic

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J Gen Physiol. 1979 Aug;74(2):187-212 PubMed

Biosci Rep. 1981 Mar;1(3):183-96 PubMed

J Fluoresc. 2004 Nov;14(6):739-43 PubMed

Eur Biophys J. 1991;19(4):183-8 PubMed

Biochemistry. 1969 Jan;8(1):322-34 PubMed

Biochim Biophys Acta. 1996 Jun 13;1282(1):101-6 PubMed

Yeast. 1998 Sep 30;14(13):1189-97 PubMed

Methods Cell Biol. 1994;41:121-33 PubMed

Biochemistry. 1974 Jul 30;13(16):3315-30 PubMed

Am J Physiol. 1993 Feb;264(2 Pt 1):C376-82 PubMed

Biochem J. 1985 Feb 1;225(3):815-9 PubMed

Biochim Biophys Acta. 1994 Dec 30;1196(2):181-90 PubMed

Rev Physiol Biochem Pharmacol. 1978;83:35-88 PubMed

Biophys J. 1993 Dec;65(6):2396-407 PubMed

Methods Cell Biol. 1989;30:193-218 PubMed

J Physiol. 1974 Jun;239(3):519-52 PubMed

Exp Cell Res. 1996 Jan 10;222(1):84-94 PubMed

Biochim Biophys Acta. 1990 Mar 15;1016(1):1-28 PubMed

J Physiol. 1976 Dec;263(2):287-319 PubMed

Biophys J. 1988 May;53(5):785-94 PubMed

J Bioenerg Biomembr. 2010 Oct;42(5):419-32 PubMed

Biochemistry (Mosc). 2005 Feb;70(2):222-30 PubMed

FEMS Yeast Res. 2006 Nov;6(7):1039-46 PubMed

J Fluoresc. 2010 Nov;20(6):1139-57 PubMed

J Bioenerg Biomembr. 1985 Jun;17(3):175-82 PubMed

J Photochem Photobiol B. 1995 Apr;28(1):93-9 PubMed

Biochim Biophys Acta. 1978 Sep 22;512(2):284-95 PubMed

J Membr Biol. 1976 Jun 30;27(4):317-34 PubMed

Stem Cell Rev Rep. 2009 Sep;5(3):231-46 PubMed

Folia Microbiol (Praha). 2000;45(3):225-9 PubMed

Anal Biochem. 2006 Jun 1;353(1):37-42 PubMed

J Photochem Photobiol B. 1996 Apr;33(2):101-24 PubMed

J Cell Comp Physiol. 1949 Dec;34(3):383-96 PubMed

Annu Rev Biophys Bioeng. 1979;8:47-68 PubMed

J Biol Chem. 2007 Jun 15;282(24):17563-7 PubMed

FEMS Yeast Res. 2008 Jun;8(4):622-30 PubMed

J Biol Chem. 2009 Jan 30;284(5):2795-2802 PubMed

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