I stained CD8 T cells with JC-1 (30min at 37°C) and acquired by FACS in FITC and PE channel immediately after. The CD8 that I used were activated for 4 days and then cultured for another 6 days. I attached a pdf with an example of my data (1st graph), cells treated with FCCP (second graph) and unstained cells (3rd graph).

I didn't see any papers that are doing this staining on mouse CD8 cells and so I don't know if the profile of my staining is correct. If I compare my staining with the images shown by companies, the separation between their 2 populations (healthy vs unhealthy) seems much more clear than mine.

Is there someone who has experience with JC-1 and could help with the gating and interpretation?