FlowJo [STAIN INDEX- difficult stains]

By popular demand- in this video we look at how to calculate stain index in FlowJo on more challenging stains that have more spread.
Looking for more FlowJo content? Let us know in the comments what other FlowJo content you would like covered!

[@becbrown8601]

Hi! My tables are just coming up with n/a and I’m confused… doing exactly what you’re showing, I don’t know why it’s happening! Thanks ☺️

[@ajarieger_flow]

Hi! Are you using the StainIndex plugin? If so, make sure you are selecting the correct parameter from the list, as well as the gates you are using (or autogate, if that's what you're going with). I've not seen n/a come up when this is done. If you're following those steps and still getting the n/a let me know!

[@石一-n4b]

love this video

[@thomson90tan]

So in your case for CD11c will you recommend to use 1:50 or 1:800 antibody dilution for your real experiment ?

[@ajarieger_flow]

Sorry I've gotten a little behind on my comments! Like any titration, you would want to look at all values (not just the high and low) and if I remember this experiment correctly, I think the best was 1:200

[@tgillson3093]

Hi, can you titrate the whole panel together, or must you titrate only one channel at a time? as you can imagine 21 colours by 5 concentrations is a lot of individual samples to run!

[@ajarieger_flow]

Generally you’ll want to titration one at a time so that (1) you don’t need to deal with compensation and (2) you know what a single antibody signal looks like so you know what you should get in your full panel (important in assessing if your panel is working correctly)

[@ajarieger_flow]

@@tgillson3093 your other comment about titrating vs voltrating and what to do first seems to have disappeared… I titrate first using somewhat optimized settings (set background and make sure positive is on scale). Once I know my best concentration, I go into voltration to see if I can further optimize my signal