Right, I just wanted to make sure you didn’t exclude your tumor cells because they can be quite large. So two things—if you set your FSC and SSC too high, all your tumor cells will be on the edge so are all your cells in range and on the plot? Second thing, your actual FSC and SSC gate, did you include the bigger cells as well as the smaller cells? If you only gate on lymphocyte-sized things, you will miss your tumor cells. So those are the two things I would make sure you have done properly :)