Cell proliferation was detected using CytoFLEX flow cytometer + fluorescent dye CFSE

The fluorescent dye CFSE (CFDA-SE) is a novel dye that can fluorescently label living cells and can label living cells. The basic principle is as follows: CFSE can easily penetrate the cell membrane, covalently bind to intracellular proteins in living cells, and release green fluorescence after hydrolysis. In the process of cell division and proliferation, its fluorescence intensity decreases with the division of cells, and the labeled fluorescence can be evenly distributed to the two daughter cells, so its fluorescence intensity is half that of the parental cells. According to this characteristic, it Can be used to detect cell proliferation, cell cycle estimation and cell division.

Materials and Methods
Reagent

2. Sample preparation

Data analysis <br>Cleavage lymphocytes on FSC/SSC, use FSC-W/FSC-A to exclude adherents, CD19-positive cells in CFSE/CD19, and lymphocytes and CD19-positive using CFSE single-sense Proliferation of (B cells).

result
Control group:

test group:

The results showed that the proliferation efficiency of CD19-positive cells after 3 days of CpG stimulation was significantly higher than that of the control group (23.44%: 18, 70%).
CytoFLEX can clearly distinguish the different algebras of lymphocyte proliferation. From the figure, there are at least 5 generations of cells, and the CD19 positive-positive differentiation is also very obvious, reflecting the superior sensitivity and resolution of the instrument.

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