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Alright! Welcome back to another FlowJo lesson here on my YouTube channel. Today Ive had more requests to look at stain index. Seems to be a hot topic! But with some more challenging stains. So I have two other examples to show you how I would gate a more challenging stain in order to do a stain index calculation. So first were going to look at an Ly6G stain. So if I open up the first plot you can see on my histogram I have a very small positive population and a really wide negative population. So it can sometimes be harder to tell which is which, so in this case I would actually change this to be a dot plot and do my gating this way. So in this case, I would gate my CD11b positives over here or sorry Ly6G positives and over here would be my Ly6G negatives and then doing this I could then go through into workspace, plugins, and my stain index and in this case again select my Ly6G and use the gates that I set up. So my positive gate, my negative gate, and cli