Comparison of different combinations of antibodies and labeled fluorescein in the detection of lymphocyte subsets by flow cytometry

Flow cytometry is a classical method for analyzing human peripheral blood lymphocyte subsets. This study aims to explore a new combination of antibody and labeled fluorescein for detecting lymphocyte subsets by comparing the effects of different combinations of antibody and labeled fluorescein in flow cytometry. We conducted a prospective study and enrolled 362 healthy patients undergoing physical examination in the medical examination center of the third hospital of hebei medical university. Venous blood was drawn from volunteers at the same time in the morning and divided into 3 tubes (Tube A, Tube B and Tube C). T lymphocytes were detected by 3-colors method (CD4-FITC/CD8-PE/CD3-PC5) in Tube A, B lymphocytes were detected by 2-colors method (CD19-FITC/CD3-PE) in Tube B, and T lymphocytes and B lymphocytes were detected by 4-colors method (CD4-FITC/CD8-PE/CD3-PC5/CD19-FITC) in Tube C. The repeatability and accuracy of the test scheme for Tube C shall not be inferior to that of Tube A and Tube B. There were no significant difference in the results of CD3 + and CD4(+/)CD8 + between Tube A and C, as well as in the results of CD3 + and CD19( + )between Tube B and C. Pearson correlation analysis showed that the test results of a and C and B and C were highly correlated. The 4-colors method (CD4-FITC/CD8-PE/CD3-PC5/CD19-FITC) can detect T lymphocytes and B lymphocytes at the same time, reduce the use of fluorescence channels and save the detection cost, which is worthy of recommendation.