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What should be done if red blood cells (RBCs) or debris are abundant in the cell suspension?
2026-06-23
Answer:
Excess RBCs and debris should be removed prior to loading to improve data quality and reduce background noise.
Recommended approaches:
- RBC lysis:
Add RBC lysis buffer at a 1:10 ratio and incubate for 3–5 minutes at room temperature, followed by centrifugation.
Avoid over-lysis, as prolonged exposure may damage nucleated cells. - Density gradient centrifugation:
Use reagents such as Ficoll or Lymphoprep to remove RBCs and dead cells. - Low-speed centrifugation:
Centrifuge at 50 × g for 3 minutes at 4°C.
Debris and RBCs remain in the supernatant, while target cells pellet.
For debris-rich samples:
- Add 0.04% BSA to the resuspension buffer to reduce aggregation and adhesion
Handle samples gently to avoid introducing additional debris
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