UNIT 9.3 Flow Cytometric Measurement of Intracellular pH
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- 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.