AIM: To explore the application value of vital dye CFDA-SE to study of lymphocytic proliferation. METHODS: CFDA-SE staining, fluorescence antibody labeling, flow cytometry and related software were used to detect the fluorescence intensity and analysis the proliferation kinetics of lymphocytes and their subsets after stimulation with polyclonal stimulators. RESULTS: Lymphocytes divided after stimulation of PDB+ionomycin or ConA for 48 h, manifesting the serial halving of fluorescence intensity. CsA inhibited the proliferative effect of ConA on lymphocytes and no CFSE fluorescence halving were seen. Proliferation of CD4(+) T cells and CD8(+) T cells were asynchronous after ConA stimulation for 48 h, which became more obvious at the time of 72 h. Proliferation-related indexs got ten from ModFit(TM) software showed that the proliferative effect of ConA on CD8(+) T cells was stronger than that on CD4(+) T cells. CONCLUSION: CFDA-SE staining combined with fluorescent antibody labeling and cytometry were powerful tools for analysis of lymphocytic proliferation.
AIM: To explore the application value of vital dye CFDA-SE to study of lymphocytic proliferation. METHODS:CFDA-SE staining, fluorescence antibody labeling, flow cytometry and related software were used to detect the fluorescence intensity and analysis the proliferation kinetics of lymphocytes and their subsets after stimulation with polyclonal stimulators. RESULTS: Lymphocytes divided after stimulation of PDB+ionomycin or ConA for 48 h, manifesting the serial halving of fluorescence intensity. CsA inhibited the proliferative effect of ConA on lymphocytes and no CFSE fluorescence halving were seen. Proliferation of CD4(+) T cells and CD8(+) T cells were asynchronous after ConA stimulation for 48 h, which became more obvious at the time of 72 h. Proliferation-related indexs got ten from ModFit(TM) software showed that the proliferative effect of ConA on CD8(+) T cells was stronger than that on CD4(+) T cells. CONCLUSION:CFDA-SE staining combined with fluorescent antibody labeling and cytometry were powerful tools for analysis of lymphocytic proliferation.