Procedure
1. Collect human peripheral blood samples into sodium citrate anticoagulant tubes.
2. Add 5 μL of fresh whole blood, 1 Test of flow cytometry antibody, and 200 μL of PBS into the centrifuge tube. Mix thoroughly and incubate at room temperature in the dark for 30 min.
3. Add 1 mL PBS to the centrifuge tube, gently vortex to mix, and proceed immediately to flow cytometric analysis.
Figure 1. Detection results of platelet marker CD61 in human peripheral blood
Precautions
1. Preferred anticoagulants: Citrate-based tubes (e.g., ACD tubes) or CTAD tubes. Avoid heparin and EDTA (EDTA induces platelet activation).
2. Use a large-bore needle (e.g., 18G) for blood collection to minimize mechanical activation.
3. Store collected samples at room temperature (15–25 °C). Temperatures below 4 °C can alter platelet morphology.
4. Time sensitivity: Perform analysis within 3 hours of collection. Delays can significantly reduce the expression of platelet-specific markers (e.g., CD62P).
5. Red blood cell lysis is not required.
6. Platelet FSC/SSC values are lower than those of red blood cells; adjust the flow cytometer threshold accordingly before acquisition.
7. Using logarithmic scales for both FSC and SSC facilitates platelet gating.
