The invention discloses a simple PBMC (peripheral blood mononuclear cell) separation method for flow cytometry detection. The method comprises the following steps: 1), taking whole blood, adding a separation solution, performing mixing, and performing standing; 2), taking the supernatant obtained in the step 1), and storing the supernatant at a temperature of 2-8 DEG C for 1-2 days to obtain a PBMC suspension; or taking the supernatant obtained in the step 1), performing centrifuging, taking lower cells, adding a preservation solution for resuspending, and performing cryopreserving for 180-360days at a temperature of -150 DEG C to obtain the cryopreserved PBMC. Compared with a common method, the method for separating PBMC has the advantages that the blood sample treatment time is greatlysaved, the operation steps are simplified, the dependence on instruments such as a centrifugal machine is reduced, and the method can be used for timely treating blood samples in wards, clinics and accident places, can be used for detecting CD4 and CD8 by flow cytometry, and has practical popularization and application values.