In some papers they sorted the cells after intracellular staining for intracellular marker such as FOXP3 (as Treg marker). But for intracellular staining, they need to fix and permeablise the cells and we know by doing so cells will NOT be viable for functional Assays such as co-culture etc. So, what method they are using for keeping cell viable and properly functional for downstream work post sorting? If possible please give references as well.
Thanks
You should check the papers you mentioned.
To my knowledge and as you said, intracellular stained cells are not alive. The sorted cells can be used for RNA/protein analysis but not for further culture.
Hi,
the surface marker CD25 and CD127 is used for Treg sorting for functional studies. as far as immunology is concerned fixed cells can still play a limited but certain role with respect to recognition by surface proteins only.
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