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Fluorimetry of mitochondria in cells vitally stained with DASPMI or rhodamine 6 GO
Author(s) -
BereiterHahn Jürgen,
Seipel KarlHeinz,
Vöth Monika,
Ploem Johan S.
Publication year - 1983
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.290010306
Subject(s) - fluorescence , rhodamine 123 , mitochondrion , antimycin a , oligomycin , biophysics , oxidative phosphorylation , cytoplasm , rhodamine , chemistry , in vivo , staining , photochemistry , biochemistry , biology , physics , microbiology and biotechnology , atpase , quantum mechanics , multiple drug resistance , enzyme , antibiotics , genetics
The fluorescent dyes DASPMI and rhodamine 6 GO specifically stain mitochondria in living cells. Dye concentrations from 10 −8 to 5 × 10 −6 mole l −1 can be used. Excitation and emission spectra, and quantum efficiency of DASPMI depend on solvent characteristics. Spectra of mitochondria in living cells correspond to those in phospholipids (excitation around 470 nm, emission 560–570 nm). Fluorescence intensity of DASPMI is a measure for the energization of mitochondria, as revealed by in vitro studies. In living cells uptake of the dye is strongly influenced by inhibitors of oxidative phosphorylation (i.e. by oligomycin, FCCP). Distribution of fluorescence intensity indicates an intracellular gradient in energy load of endothelial cells. Single mitochondria exhibit oscillations in fluorescence. Mitochondria loaded with DASPMI release the dye suddenly into the cytoplasm on treatment with FCCP. Cyanide and antimycin however, do not diminish fluorescence in vivo under optimal nutritional conditions, while they do so in mitochondrial suspension, indicating different mitochondrial behaviour in vivo and in suspension.