Two or more cell populations in Flow cytometry
Explore possible causes and solutions for seeing two or more cell populations when there should only be one in Flow cytometry.
Why are there 2 or more cell populations observed when there should be just one?
Possible causes
Solution
More than one cell population present expressing target protein
- Check expected expression levels from the cell types contained in the sample and ensure adequate cell separation if necessary.
Cell doublets present.
Doublets of cells will show as a second cell population at approximately twice the fluorescence intensity on the plot.
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Mix cells gently before staining and again before running on the cytometer using a pipette.
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Cells can also be sieved or filtered to remove clumps (30 μl Nylon Mesh).