what's the cell subset on the left of cd4 t cell, after i purified cd4 t cell from mouse lymphnode and activate with anti-cd3/28? x-FSC y-SSC
they are dead cells and dying cells. Sometimes too much activation by CD3/CD28 cause to die.
Yup, they're dead. Try using a viability dye to label them and you'll be able to exclude them from your analyses.
Try the fixable viability dyes from Life Tech - I've used several over the years and they're excellent -
https://www.lifetechnologies.com/uk/en/home/life-science/cell-analysis/flow-cytometry/cell-health-and-viability-assays-for-flow-cytometry/cell-viability-assays-for-flow-cytometry/flow-fixable-viability-dyes.html
there are three subsets. which one are you referring to? My guess will be the left one will be debris but that will left us with the middle and the right subsets. what are the middle and the right subsets then?
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