Bereiter-Hahn J, Seipel K H, Vöth M, Ploem J S
Cell Biochem Funct. 1983 Oct;1(3):147-55. doi: 10.1002/cbf.290010306.
The fluorescent dyes DASPMI and rhodamine 6 GO specifically stain mitochondria in living cells. Dye concentrations from 10(-8) to 5 X 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.
荧光染料DASPMI和罗丹明6G可特异性地对活细胞中的线粒体进行染色。染料浓度范围为10^(-8)至5×10^(-6)摩尔/升。DASPMI的激发光谱、发射光谱以及量子效率取决于溶剂特性。活细胞中线粒体的光谱与磷脂中的光谱相对应（激发波长约为470纳米，发射波长为560 - 570纳米）。体外研究表明，DASPMI的荧光强度是衡量线粒体能量化的指标。在活细胞中，染料的摄取受到氧化磷酸化抑制剂（如寡霉素、FCCP）的强烈影响。荧光强度分布表明内皮细胞能量负荷存在细胞内梯度。单个线粒体的荧光呈现振荡现象。用FCCP处理时，加载有DASPMI的线粒体将染料突然释放到细胞质中。然而，在最佳营养条件下，氰化物和抗霉素在体内并不会降低荧光，而在悬浮的线粒体中则会降低，这表明线粒体在体内和悬浮状态下的行为有所不同。
