Comparative study of whole blood lysis reagents for analysis of immunocytes in peripheral blood of mice by flow cytometry.
- Author:
Wei CUI
1
;
Sa LIU
;
Lun CAI
;
Yu-Lin LI
;
Cong-Cong ZHANG
;
Shu-Lan QIU
Author Information
1. Capital Medical University, Beijing 100029, China. cuiwei1117@sina.com
- Publication Type:Journal Article
- MeSH:
Animals;
Erythrocyte Count;
Erythrocytes;
immunology;
metabolism;
Flow Cytometry;
instrumentation;
methods;
Immune System;
immunology;
Indicators and Reagents;
analysis;
Mice;
Mice, Inbred C57BL
- From:
Journal of Experimental Hematology
2011;19(2):491-495
- CountryChina
- Language:Chinese
-
Abstract:
This study was purposed to investigate the efficacy of different whole flow lysis reagents for lysis of red blood cells in flow cytometric analysis. The expression of immunocytes was detected by flow cytometry after lysis of red blood cells using commercial reagents (Optilyse C, FACS Lysing Solution) and self-made red blood cell lysis reagents (RBC Lysis Buffer), the detection results were analyzed comparatively. The results showed that there was no significant difference in the percentage of CD3e(+), CD3e(+)CD4(+), CD3e(+)CD8a(+), CD3e(-)CD19(+), CD3e(-)NK1.1(+) and Gr-1(+) cells between 3 different lysis reagent groups. However OptiLyse C solution was suitable to Gr-1(+) cell detection, but did not suit to Foxp3(+) Treg detection. The self-made RBC Lysis Buffer and FACS Lysing Solution were suited to Foxp3(+) Treg detection. It is concluded that the use of self-made RBC Lysis Buffer for flow cytometry can get the lysis efficiency of commercially available lysis solutions when samples are prepared in accordance with standardized procedure. The self-made RBC Lysis Buffer not only can satisfy experimental requirements, but also can reduce the experimental costs.
