A method for preparing lymphocyte sample for flow cytometry analysis

A method for preparing flow cytometric sample in immune cells for flow cytometry, comprising the steps of:

• 1) separating plasma and blood precipitation by centrifuging in vitro anticoagulant blood samples to obtain blood cell precipitate;
• 2) mixing the blood cell precipitate with PBS solution to obtain a blood cell dilution;
• 3) adding the blood cell dilution into a centrifuge tube containing the lymphocyte separation solution having a volume equal to that of the blood cell dilution, centrifuging, and discarding plasma to obtain buffy coat cells;
• 4) adding the buffy coat cells to a centrifuge tube, adding PBS solution under stirring, centrifuging, discarding supernatant, and then adding PBS solution to adjust the cell concentration to provide lymphocyte suspension having a concentration of 1×105/ml-1×108/ml;
• 5) adding the lymphocyte suspension into a flow tube, then adding flow cytometric antibody to label lymphocyte subpopulations, mixing evenly, and incubating at 2-8° C. without light;
• 6) adding PBS solution to the flow tube, mixing, and centrifuging to remove unbound antibody;
• 7) completion of samples;
• {circle around (1)} If a detection on a flow cytometer is performed immediately, the supernatant is discarded and the precipitate is mixed with PBS-EDTA solution at 2-8° C. for flow cytometric detection;
• or
• {circle around (2)} If the detection cannot be performed immediately, formalin is added, mixed, and left to stand at 2-8° C. without light;
• and before detection, each of the tubes is added with PBS solution, centrifuged to discard the supernatant, and evenly mixed with PBS-EDTA solution at 2-8° C. for flow cytometric detection.