Hi.
I am doing flow cytometry of endometrial cells (read epithelial and fibroblast). The antibodies I need to use are both intracellular. In the same time I want to get only life cells.
Thus I am using the fixable life/dead reagent before do the permeabilization, to filter only live cells of my samples.
however when I read on flow cytometry, right after the permeabilization all my life population disappear.
I am attaching two pictures, One before, and other right after the permeabilization. You can see there is no life cells to gate it.
Do you guys know any explanation for it?
Thanks
1. I can see only one picture here. If you are saying two graph as two picture then in second this show only your selected population is negative for both RL1-A and BL1-A.
2. No need to filter the cells you opt out by gating.
3. And Live dead should be selected in FSC vs SSC plot.
Hey Ankesh. Thanks for your prompt response. I made a picture format where you can find my both graphs. left is before perm, how much life cells I had and right is after permed, following the manufactures instructions
The reagent I am using to color those live cells is called:
LIVE/DEAD Fixable Dead Cell Stains from Fisher Scientific (cat. L23101). Its is color RL3 in my flow cytometer. it was suppose to keep the same thing before and after perm, but this is not happen with me
Do yo know if is possible that some interactions are happen with this ?
the picture of the graphs is attached bellow
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