Hi..could someone briefly explain how to correctly eliminate dead cells using on Forward Scatter and side scatter ?
if u know about your cell of interest cell size and internal cellular complexity then u can eliminate the dead cells.
Or else u can stain the cell of interest and the run on the machine. While running u can apply gate on the different clusters u get in the FSC vs SSC plot and show the gated clusters in the flourescent plots and see were it falls.
I would not recommend using FSC and SSC for live/dead determination. At best you may be able to enrich for live cells, but not exclude them. A proper viability dye must be used, such as PI, 7AAD, or DAP (if using live cells) or Fixable Viability Dyes like those available from Life Technologies (if using fixed cells).
I would also use PI, wichi as Debbie points out will not penetrate the cell membrane. However, be careful to add the PI just when you are going to put the sample on the FACS (if you leave it for a long time all cells will become positive....). We usually add it to each sample before it gets into the FACS. with this you can gate back to your forward and side scatter plots to see if you can separete the population there.
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