UNIT 9.3 Flow Cytometric Measurement of Intracellular pH

  1. Sue Chow,
  2. David Hedley

Published Online: 1 MAY 2001

DOI: 10.1002/0471142956.cy0903s14

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Chow, S. and Hedley, D. 2001. Flow Cytometric Measurement of Intracellular pH. Current Protocols in Cytometry. 00:9.3:9.3.1–9.3.10.

Author Information

  1. Ontario Cancer Institute and Princess Margaret Hospital, Toronto, Ontario, Canada

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 2000


A number of fundamentally important biological processes, such as cell signaling and the initiation of mitosis, are accompanied by a change in intracellular pH. Flow cytometric measurement of pH is a generally straightforward procedure that can be done with any instrument equipped with a 488-nm argon laser. The overall approach is similar to that for calcium: generation of a calibration curve by imparting known changes in pH and interpolation of the test sample pH. This unit presents the traditional calibration method using high-potassium buffers and the proton ionophore nigericin and a more recently developed technique, the pseudo null method, which involves resuspension of cells in defined mixtures of weak acids and weak bases.