Flow cytometry detection method for mouse thymus or spleen T-lymphocyte subsets
A flow cytometry and lymphocyte technology, which is applied in the field of flow cytometry detection of mouse thymus or spleen T lymphocyte subsets, can solve the problems of large fluctuation and instability in the detection results, and achieve easy control and avoid errors. , the effect of simple operation steps
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[0051] Step 1. Obtain the spleen or thymus tissue of the subject mouse and prepare a single cell suspension
[0052] According to a certain dose level, cyclophosphamide was injected intraperitoneally to the experimental mice for one week to establish an immunosuppressive model. The mice that were successfully modeled were sacrificed by cervical dislocation and killed immediately, the spleen or thymus tissue was taken out, the blood was blotted with filter paper, the surrounding connective tissue was trimmed with ophthalmic scissors, and stored in PBS at 4°C;
[0053] Take the preserved sample, put it in a watch glass, cut it repeatedly with ophthalmic scissors until the tissue becomes muddy, add an appropriate amount of PBS solution, and mix well; then filter it with a 300-mesh nylon mesh to obtain the filtrate; at a speed of 600-1000r / min Centrifuge the filtrate for 5 minutes, discard the supernatant; then add 1mL PBS solution, vortex to mix, centrifuge again, discard the sup...
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