Miyata H, Hayashi H, Suzuki S, Noda N, Kobayashi A, Fujiwake H, Hirano M, Yamazaki N
Third Department of Internal Medicine, Hamamatsu University School of Medicine, Japan.
Biochem Biophys Res Commun. 1989 Aug 30;163(1):500-5. doi: 10.1016/0006-291x(89)92165-7.
Isolated rat heart myocytes were loaded with both the Ca2+ sensitive fluorescent probe fura-2/AM and the fluorescent pH indicator 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF/AM). Changes in [Ca2+]i and pHi were measured simultaneously using digitized video fluorescence microscopy. In measurement of [Ca2+]i and pHi, the ratios of dual-loaded cells were not different from single-loaded cells. Using this method, [Ca2+]i and pHi in myocytes were 48 +/- 7 nM and 7.17 +/- 0.05. It is concluded that [Ca2+]i and pHi could be measured simultaneously in isolated myocyte using dual-loading of fura-2 and BCECF.
将分离的大鼠心肌细胞同时加载钙离子敏感荧光探针fura-2/AM和荧光pH指示剂2,7-双羧乙基-5(6)-羧基荧光素（BCECF/AM）。使用数字化视频荧光显微镜同时测量细胞内钙离子浓度（[Ca2+]i）和细胞内pH值（pHi）的变化。在测量[Ca2+]i和pHi时，双加载细胞的比率与单加载细胞没有差异。使用这种方法，心肌细胞中的[Ca2+]i和pHi分别为48±7 nM和7.17±0.05。结论是，使用fura-2和BCECF双加载可以在分离的心肌细胞中同时测量[Ca2+]i和pHi。
