What should be checked if the number of detected cells is much lower than expected (excluding clogging)?

2026-06-23

Answer:
If clogging is excluded, the following factors should be evaluated:

Stepwise troubleshooting:

  • Sample quality: 
    • Viability >90%? 
    • Nucleated cell proportion >70%? 
  • Cell counter images:
    Check for debris or RBC contamination (which may artificially inflate counts) 
  • UMI distribution:
    If separation between true cells and background is unclear, adjust the cutoff threshold (e.g., 200–500 UMI) 

Biological factors:
Certain cell types (e.g., neutrophils, platelets) naturally yield fewer detectable transcripts

 

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