This experiment is based on "Color Atlas of Practical Flow Cytometry", edited by Shukui Wang and Zhenying Zhou.

Operation method

Peripheral Blood Immune Cell Assay

Materials and Instruments

Venous whole blood
Heparin Mouse Anti-Human Targeted Monoclonal Antibody Erythrocyte Lysate Saline
FCM measuring tubes

Move

1. Take 100 ul of 1000 U/ml heparin anticoagulated venous whole blood and put it into the FCM measuring tube.

2. Direct labeling method: add 10 ul of fluorescently labeled murine anti-human target monoclonal antibody reagent, mix well and react for 30 min in the refrigerator at 4 ℃.

3. Indirect labeling method: add 10 ul of non-fluorescently labeled murine anti-human target monoclonal antibody reagent, mix well and react for 30 min. 4. Add 2 ml of lysed red cell solution, mix well and react for 5~10 min in the refrigerator at 4 ℃. 5. Add 10 ul of mouse anti-mouse monoclonal antibody without fluorescent marker, mix thoroughly, and react for 30 min.

4. Add 2 ml of erythrocyte solution, mix thoroughly, and react for 5~10 min in the refrigerator at 4℃.

5. Centrifuge at 1500 r/min for 5 min, and then remove the supernatant.

6. Wash the cells with physiological saline for one time, and then collect the cells, and then test them on the machine.

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