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Dual loading of the fluorescent indicator fura-2 and 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) in isolated myocytes

1989; Elsevier BV; Volume: 163; Issue: 1 Linguagem: Inglês

10.1016/0006-291x(89)92165-7

1090-2104

Haruo Miyata, Hideharu Hayashi, Shingo Suzuki, Naohisa Noda, Akira Kobayashi, Hideshi Fujiwake, Masahiko Hirano, Noboru Yamazaki,

Electrochemical Analysis and Applications

Isolated rat heart myocytes were loaded with both the Ca2+ sensitive fluorescent probe fura-2AM 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.