Membrane cell permeabilisation with saponin and multiparametric analysis by flow cytometry
Article first published online: 8 MAR 2005
Copyright © 1991 Wiley-Liss, Inc.
Volume 12, Issue 6, pages 550–558, 1991
How to Cite
Jacob, M. C., Favre, M. and Bensa, J.-C. (1991), Membrane cell permeabilisation with saponin and multiparametric analysis by flow cytometry. Cytometry, 12: 550–558. doi: 10.1002/cyto.990120612
- Issue published online: 21 JUN 2005
- Article first published online: 8 MAR 2005
- Manuscript Accepted: 24 MAR 1991
- Manuscript Received: 22 NOV 1989
- intracellular antigen;
- multiparametric analysis;
- flow cytometry
Saponin, a detergent like molecule, can permeabilise cell membranes without destroying them, and thus can be used for the detection of intracellular antigens on intact cells with a flow cytometer.
First experiments were reported that demonstrated the detection of intracytoplasmic antigens such as intermediate filaments and CD3 in T acute lymphoblastic leukemia (ALL). Further experiments were also performed to prove that intranuclear structures were equally accessible: dyes such as propidium iodide (PI) and monoclonal antibodies (mAb) such as Ki67 could penetrate the nucleus and lead to the analysis of DNA content and to the discrimination between the different cell cycle phases (G0, G1, S, G2-M).
This rapid and sensitive method retained sufficient integrity of cells being treated to enable differentiation of cell types on the basis of morphology. Furthermore, it did not alter membrane expression of most antigens. Therefore, it was of particular interest for multiparametric analysis, especially for simultaneous study of membrane and intracellular structures.