I have Treg (CD4+YFP+) cells and Tcon (CD4+) cells in coculture. The proliferation marker is Cell Trace Violet. Post harvesting, I stain my samples for live dead staining with blue fluorescent reactive and then surface staining with CD4. I use flowjo for analysis. Can I directly gate around the live cell population to see dye dilution as on Tcon cells are stained with CTV or I should specifically gate around the CD+YFP- cells and then gate this population to check dye dilution?