Post Sat Jun 11, 2022 1:08 am

Re: Double CD8 population

Hi Chad

Few things that can happen - saying this as I saw this double pop few times:

I suggest to run the same sample using mono marker. In samples we had that issue we got out this population by gating CD3 *after* monos
so cleanup->single->alive->CD45+->CD19vsCD33(orCD14)->CD3vsCD56, once you clean your CD3s from monos you we lost this pop. Gating on cd45+ Cd3- wasnt enough as we did observe importnat unspecific staining of monos with cd8. I highly recommend to gate your population of interest after the most precise cleanup possible, so if you focus on T, clean it first from Bs and myeloids etc

Also another possibility - question - is your Ab from fluidigm or in-house conjugation. We had issues with conjugation stability for long storage, but it was rather weaker signal than double pop

Another eventuality - we saw in some pts gdT and MAIT cells coexpressing CD8

Jo :ugeek: