A microfluorometric method for simultaneous measurement of changes in cytosolic free calcium concentration and pH in single cardiac myocytes.
We describe a new system to measure changes in cytosolic free Ca concentration ([Ca2+]i) and pH (pHi) simultaneously using the dyes fura-2 and BCECF, and a random access camera with computer-operated synchronously-changing excitation and emission filters. Fluorescence intensities for fura-2 and BCECF were recorded from rat cardiac myocytes and calibrated in vivo using multiple-point determinations. [Ca2+]i and pHi were 88.6 +/- 32.8 nM and 7.16 +/- 0.12, respectively.
Department of Physiology, Yamagata University School of Medicine, Japan.
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Shibuya
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Matsuyama
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Tanaka
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Doi
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003592:Cytophotometry / Q000379:methods*; D003600:Cytosol / Q000378:metabolism; D005452:Fluoresceins; D016257:Fura-2; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D005452:Fluoresceins; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D002118:Calcium; D016257:Fura-2