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

Raul Martínez-Zaguilán, Mark W. Gurulé, Ron Lynch

Research output: Contribution to journalArticle

40 Citations (Scopus)

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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume270
Issue number5 39-5
StatePublished - May 1996

Fingerprint

Fura-2
Insulin-Secreting Cells
Microscopy
Microscopic examination
Ions
Insulin
Imaging techniques
Glucose
Spectrographs
Acidification
CCD cameras
Insulinoma
Microscopes
Cricetinae
Homeostasis
Light
Equipment and Supplies
seminaphthorhodaminefluoride

Keywords

  • Fluorescence
  • Fura 2
  • HIT-T15
  • Microfluorometry
  • SNARF-1

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology
  • Physiology (medical)

Cite this

Simultaneous measurement of intracellular pH and Ca2+ in insulin-secreting cells by spectral imaging microscopy. / Martínez-Zaguilán, Raul; Gurulé, Mark W.; Lynch, Ron.

In: American Journal of Physiology - Cell Physiology, Vol. 270, No. 5 39-5, 05.1996.

Research output: Contribution to journalArticle

@article{e678c8ef38f4430cbe9966b9a7b39ec2,
title = "Simultaneous measurement of intracellular pH and Ca2+ in insulin-secreting cells by spectral imaging microscopy",
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.",
keywords = "Fluorescence, Fura 2, HIT-T15, Microfluorometry, SNARF-1",
author = "Raul Mart{\'i}nez-Zaguil{\'a}n and Gurul{\'e}, {Mark W.} and Ron Lynch",
year = "1996",
month = "5",
language = "English (US)",
volume = "270",
journal = "American Journal of Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "5 39-5",

}

TY - JOUR

T1 - Simultaneous measurement of intracellular pH and Ca2+ in insulin-secreting cells by spectral imaging microscopy

AU - Martínez-Zaguilán, Raul

AU - Gurulé, Mark W.

AU - Lynch, Ron

PY - 1996/5

Y1 - 1996/5

N2 - 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.

AB - 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.

KW - Fluorescence

KW - Fura 2

KW - HIT-T15

KW - Microfluorometry

KW - SNARF-1

UR - http://www.scopus.com/inward/record.url?scp=0029896495&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029896495&partnerID=8YFLogxK

M3 - Article

C2 - 8967445

AN - SCOPUS:0029896495

VL - 270

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0363-6143

IS - 5 39-5

ER -