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Flow cytometry is often used for detection of various proteins/antigens in blood samples, particularly for immunophenotyping. Analysis of the results is usually much simpler and more accurate when the red blood cells are not included in the sample and have been lysed.
There are several commercially available red blood cell lysis kits available (manufacturers kit protocol should be followed).
Several detergents have been reported to affect the detection of surface and internal antigens of red blood cells. In the following reference the impact of different detergents and detergent concentrations on red blood cell lysis and on whole blood cell resolution using light scatter was investigated.
Only three detergents, Triton X 100, NP-40 and Brij-58, showed significant red blood cell lysis after 2% formaldehyde fixation for ten minutes at room temperature. These detergents had a deleterious effect on whole blood cell light scatter at detergent concentration above 0.2%, whereas concentration below 0.1% resulted in incomplete red blood cell lysis. Incubation for 30 min with 0.1% detergent was optimal.
Adapted from: Chow S, Hedley D, Grom P, Magari R, Jacobberger JW and Shankey TV. (2005) Whole blood fixation and permeabilization protocol with red blood cell lysis for flow cytometry of intracellular phosphorylated epitopes in leukocyte subpopulations. Cytometry 67, 4–17.