Dual loading of the fluorescent indicator fura-2 and 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) in isolated myocytes.
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.
Third Department of Internal Medicine, Hamamatsu University School of Medicine, Japan.
H
H
Miyata
H
H
Hayashi
S
S
Suzuki
N
N
Noda
A
A
Kobayashi
H
H
Fujiwake
M
M
Hirano
N
N
Yamazaki
D000818:Animals; D001572:Benzofurans / Q000008:administration & dosage*; D002118:Calcium / Q000032:analysis*; D005452:Fluoresceins / Q000008:administration & dosage*; D016257:Fura-2; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008856:Microscopy, Fluorescence / Q000379:methods; D009206:Myocardium / Q000166:cytology*; D051381:Rats; D014741:Video Recording
D001572:Benzofurans; D005452:Fluoresceins; D007202:Indicators and Reagents; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D002118:Calcium; D016257:Fura-2