UNIT 5.5 Measurement of Intracellular Ions by Flow Cytometry

  1. Carl H. June1,
  2. Ryo Abe1,
  3. Peter S. Rabinovitch2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0505s14

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

June, C. H., Abe, R. and Rabinovitch, P. S. 2001. Measurement of Intracellular Ions by Flow Cytometry. Current Protocols in Immunology. 5:5.5.

Author Information

  1. 1

    Naval Medical Research Institute, Bethesda, Maryland

  2. 2

    University of Washington, Seattle, Washington

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1995

This is not the most recent version of the article. View current version (1 DEC 2004)


The recent development of a number of new fluorescent probes makes it possible to measure the concentrations of various intracellular free ions in single living cells. Among these ions are calcium, magnesium, sodium, potassium, and hydrogen (pH). This unit describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer.