Sample preparation experiments for flow cytometry by direct fluorescent staining of micro-volume whole blood

Summary

This experimental method was obtained from the official website of the Fourth Military Medical University

Operation method

Sample preparation experiments for flow cytometry by direct fluorescent staining of micro-volume whole blood

Move

Currently there are many methods to prepare whole blood cell samples and they are very mature. The commonly used method is to separate lymphocytes with lymphocyte isolation solution, and then carry out specific fluorescence staining, which is the same as the method described earlier. In the following, we mainly introduce the Q-PREP (Immunology Sample Processor), which is compatible with the flow cytometer, for the rapid preparation of micro whole blood samples.

1. Take heparin or EDTA anticoagulated whole blood (100μl/sample) and place it in a 12×75mm special plastic test tube.

2. Add 20μl of specific fluorescent monoclonal antibody to each portion, and add fluorescent labeled irrelevant monoclonal antibody to the other portion as isotype control, and then stain the blood for 15min at room temperature and protected from light.

3. Lysis of erythrocytes, stabilization and fixation of leukocytes on Q-PREP instrument, and leave for 5min.

4. Detect on flow cytometer.

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