PMID: 8967445May 1, 1996Paper

Simultaneous measurement of intracellular pH and Ca2+ in insulin-secreting cells by spectral imaging microscopy

The American Journal of Physiology
R Martinez-ZaguilanRonald M Lynch

Abstract

Described is a microscopic spectral imaging approach to monitor pH and Ca2+ simultaneously from combined spectra of multiple ion indicators. Emitted light from a cell is focused onto a grating spectrograph and spectra are imaged with a cooled charge-coupled device camera. The combined spectral output of fura 2 and SNARF-1 was analyzed to follow changes in intracellular Ca2+ concentration ([Ca2+]i) and intracellular pH (pHi) simultaneously and to correct the Ca2+ signal for concurrent changes in pHi. Responses of individual hamster insulinoma (HIT-T15) cells to effectors of ion homeostasis were heterogeneous. Treatment with NH4Cl increased pHi and transiently increased [Ca2+]i. Removal of NH4Cl induced cytosolic acidification concomitant with either no change or sustained increases in [Ca2-]i. Glucose treatment generally resulted in rapid and sustained increases in both [Ca2+]i and pHi but also heterogeneous pHi and [Ca2+]i responses. Corrections of the fura 2 signal for pH were important for following Ca2+ transitions elicited by NH4Cl but were less important for glucose-induced responses. The spectral imaging microscope provides a sensitive method for simultaneous measurements of pHi and [Ca2+]i in single cells.

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Citations

May 11, 2002·American Journal of Physiology. Heart and Circulatory Physiology·O GryshchenkoR D Nathan
Mar 10, 1998·The Journal of Physiology·T Speake, A C Elliott
Jul 10, 2001·American Journal of Physiology. Cell Physiology·H XuF K Ghishan
Feb 28, 2008·Applied Optics·C E VolinG H Bearman
Sep 3, 2013·Physiology·Shady SaadReinhard Dechant
May 18, 2011·The Journal of General Physiology·Yuanyuan Y WangEmily R Liman
May 5, 2005·Applied and Environmental Microbiology·Ryan C Hunter, Terry J Beveridge
Nov 9, 2018·American Journal of Physiology. Cell Physiology·Nathaniel J HartRonald M Lynch
Dec 31, 1997·The American Journal of Physiology·K N RichmondR M Lynch
Jul 11, 2002·American Journal of Physiology. Cell Physiology·Linda S TompkinsRonald M Lynch
Jun 9, 1998·Journal of Cellular Physiology·R Martínez-ZaguilánR J Gillies
Dec 7, 2007·Journal of Bioenergetics and Biomembranes·Souad R Sennoune, Raul Martinez-Zaguilan
May 9, 2006·American Journal of Physiology. Heart and Circulatory Physiology·J D RojasR Martínez-Zaguilán
Sep 2, 2005·American Journal of Physiology. Cell Physiology·Sergio Sánchez-ArmássRaul Martínez-Zaguilán
Jul 31, 1998·Archives of Biochemistry and Biophysics·C A HudsonR Martínez-Zaguilán

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