Open AccessSimultaneous measurement of Ca 2+ and pH by laser cytometry using fluo‐3 and SNARF‐1
Author(s) -
Atsumi Toshiko,
Sugita Kenji,
Kohno Minoru,
Takahashi Tsutomu,
Ueha Takao
Publication year - 1996
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19960601)24:2<99::aid-cyto1>3.0.co;2-c
Subject(s) - bapta , intracellular , biophysics , calcium , flow cytometry , calcium in biology , chemistry , cytometry , intracellular ph , microbiology and biotechnology , biology , biochemistry , organic chemistry
We present a new convenient method for simultaneous measurement of intracellular calcium concentration ([Ca 2+ ] i ) and intracellular pH (pH i ) using laser cytometry with a mixture of fluo‐3 (for [Ca 2+ ] i ) and SNARF‐1 (for pH i ), with iso excitation (488 nm)‐dual emission (530 nm for fluo‐3 and > 630 nm for SNARF‐1). By using this technique, we measured the changes in [Ca 2+ ] i and pH i in A‐431 human epidermoid carcinoma cells and HSG human salivary gland cells stimulated by ATP. We found that alkalization in A‐431 cell occurred with the elevation of [Ca 2+ ] i ; in contrast, alkalization in HSG cells did not occur at all, even though the elevation of [Ca 2+ ] i was observed. Using BAPTA (a chelating agent of Ca 2+ ) and amiloride (an inhibitor of the Na + /H + exchanger), we found that the elevation of pH i requires the elevation of [Ca 2+ ] i but that the elevation of [Ca 2+ ] i does not always require a rise in pH i . From our results we conclude that elevation of [Ca 2+ ] i takes precedence over the elevation of pH i in ATP‐stimulated signal transduction. © 1996 Wiley‐Liss, Inc.