Single cell and subcellular measurements of intracellular Ca2+ concentration

John G. McCarron, Marnie L. Olson, Susan Chalmers, John M. Girkin

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Increases in bulk average cytoplasmic Ca2+ concentration ([Ca2+]c) are derived from the combined activities of many Ca2+ channels. Near (<100 nm) the mouth of each of these channels the local [Ca2+]c rises and falls more quickly and reaches much greater values than occurs in the bulk cytoplasm. Even during apparently uniform, steady-state [Ca2+] increases large local inhomogeneities exist near channels. These local increases modulate processes that are sensitive to rapid and large changes in [Ca2+] but they cannot easily be visualized with conventional imaging approaches. The [Ca 2+] changes near channels can be examined using total internal reflection fluorescence microscopy (TIRF) to excite fluorophores that lie within 100 nm of the plasma membrane. TIRF is particularly powerful when combined with electrophysiology so that ion channel activity can be related simultaneously to the local subplasma membrane and bulk average [Ca2+]c. Together these techniques provide a better understanding of the local modulation and control of Ca2+ signals.

LanguageEnglish
Title of host publicationCalcium Signaling Protocols
EditorsDavid G. Lambert, Richard D. Rainbow
Pages239-251
Number of pages13
Volume937
DOIs
Publication statusPublished - 2013

Publication series

NameMethods in Molecular Biology
Volume937
ISSN (Print)10643745

Fingerprint

Fluorescence Microscopy
Electrophysiology
Ion Channels
Mouth
Cytoplasm
Cell Membrane
Membranes

Keywords

  • fluorescence imaging
  • image analysis
  • patch clamp
  • smooth muscle cell isolation
  • TIRF

Cite this

McCarron, J. G., Olson, M. L., Chalmers, S., & Girkin, J. M. (2013). Single cell and subcellular measurements of intracellular Ca2+ concentration. In D. G. Lambert, & R. D. Rainbow (Eds.), Calcium Signaling Protocols (Vol. 937, pp. 239-251). (Methods in Molecular Biology; Vol. 937). https://doi.org/10.1007/978-1-62703-086-1
McCarron, John G. ; Olson, Marnie L. ; Chalmers, Susan ; Girkin, John M. / Single cell and subcellular measurements of intracellular Ca2+ concentration. Calcium Signaling Protocols. editor / David G. Lambert ; Richard D. Rainbow. Vol. 937 2013. pp. 239-251 (Methods in Molecular Biology).
@inbook{b9d1823a8575415b9f32fe4fc90bb5d1,
title = "Single cell and subcellular measurements of intracellular Ca2+ concentration",
abstract = "Increases in bulk average cytoplasmic Ca2+ concentration ([Ca2+]c) are derived from the combined activities of many Ca2+ channels. Near (<100 nm) the mouth of each of these channels the local [Ca2+]c rises and falls more quickly and reaches much greater values than occurs in the bulk cytoplasm. Even during apparently uniform, steady-state [Ca2+] increases large local inhomogeneities exist near channels. These local increases modulate processes that are sensitive to rapid and large changes in [Ca2+] but they cannot easily be visualized with conventional imaging approaches. The [Ca 2+] changes near channels can be examined using total internal reflection fluorescence microscopy (TIRF) to excite fluorophores that lie within 100 nm of the plasma membrane. TIRF is particularly powerful when combined with electrophysiology so that ion channel activity can be related simultaneously to the local subplasma membrane and bulk average [Ca2+]c. Together these techniques provide a better understanding of the local modulation and control of Ca2+ signals.",
keywords = "fluorescence imaging, image analysis, patch clamp, smooth muscle cell isolation, TIRF",
author = "McCarron, {John G.} and Olson, {Marnie L.} and Susan Chalmers and Girkin, {John M.}",
year = "2013",
doi = "10.1007/978-1-62703-086-1",
language = "English",
isbn = "978-1-62703-086-1",
volume = "937",
series = "Methods in Molecular Biology",
pages = "239--251",
editor = "Lambert, {David G.} and Rainbow, {Richard D.}",
booktitle = "Calcium Signaling Protocols",

}

McCarron, JG, Olson, ML, Chalmers, S & Girkin, JM 2013, Single cell and subcellular measurements of intracellular Ca2+ concentration. in DG Lambert & RD Rainbow (eds), Calcium Signaling Protocols. vol. 937, Methods in Molecular Biology, vol. 937, pp. 239-251. https://doi.org/10.1007/978-1-62703-086-1

Single cell and subcellular measurements of intracellular Ca2+ concentration. / McCarron, John G.; Olson, Marnie L.; Chalmers, Susan; Girkin, John M.

Calcium Signaling Protocols. ed. / David G. Lambert; Richard D. Rainbow. Vol. 937 2013. p. 239-251 (Methods in Molecular Biology; Vol. 937).

Research output: Chapter in Book/Report/Conference proceedingChapter

TY - CHAP

T1 - Single cell and subcellular measurements of intracellular Ca2+ concentration

AU - McCarron, John G.

AU - Olson, Marnie L.

AU - Chalmers, Susan

AU - Girkin, John M.

PY - 2013

Y1 - 2013

N2 - Increases in bulk average cytoplasmic Ca2+ concentration ([Ca2+]c) are derived from the combined activities of many Ca2+ channels. Near (<100 nm) the mouth of each of these channels the local [Ca2+]c rises and falls more quickly and reaches much greater values than occurs in the bulk cytoplasm. Even during apparently uniform, steady-state [Ca2+] increases large local inhomogeneities exist near channels. These local increases modulate processes that are sensitive to rapid and large changes in [Ca2+] but they cannot easily be visualized with conventional imaging approaches. The [Ca 2+] changes near channels can be examined using total internal reflection fluorescence microscopy (TIRF) to excite fluorophores that lie within 100 nm of the plasma membrane. TIRF is particularly powerful when combined with electrophysiology so that ion channel activity can be related simultaneously to the local subplasma membrane and bulk average [Ca2+]c. Together these techniques provide a better understanding of the local modulation and control of Ca2+ signals.

AB - Increases in bulk average cytoplasmic Ca2+ concentration ([Ca2+]c) are derived from the combined activities of many Ca2+ channels. Near (<100 nm) the mouth of each of these channels the local [Ca2+]c rises and falls more quickly and reaches much greater values than occurs in the bulk cytoplasm. Even during apparently uniform, steady-state [Ca2+] increases large local inhomogeneities exist near channels. These local increases modulate processes that are sensitive to rapid and large changes in [Ca2+] but they cannot easily be visualized with conventional imaging approaches. The [Ca 2+] changes near channels can be examined using total internal reflection fluorescence microscopy (TIRF) to excite fluorophores that lie within 100 nm of the plasma membrane. TIRF is particularly powerful when combined with electrophysiology so that ion channel activity can be related simultaneously to the local subplasma membrane and bulk average [Ca2+]c. Together these techniques provide a better understanding of the local modulation and control of Ca2+ signals.

KW - fluorescence imaging

KW - image analysis

KW - patch clamp

KW - smooth muscle cell isolation

KW - TIRF

UR - http://www.scopus.com/inward/record.url?scp=84873537567&partnerID=8YFLogxK

U2 - 10.1007/978-1-62703-086-1

DO - 10.1007/978-1-62703-086-1

M3 - Chapter

SN - 978-1-62703-086-1

VL - 937

T3 - Methods in Molecular Biology

SP - 239

EP - 251

BT - Calcium Signaling Protocols

A2 - Lambert, David G.

A2 - Rainbow, Richard D.

ER -

McCarron JG, Olson ML, Chalmers S, Girkin JM. Single cell and subcellular measurements of intracellular Ca2+ concentration. In Lambert DG, Rainbow RD, editors, Calcium Signaling Protocols. Vol. 937. 2013. p. 239-251. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-086-1