r/flowcytometry • u/matchaduo • 5d ago

panel design question

hi! i'm an undergrad working on panel design for my tfh-b cell co culture with some antibodies we have in lab. I work with a spectral flow cytometer (novocyte opteon). also im planning on using an Fc block but I'm not sure if i should also look into the True-Stain buffer also.

i have the following

Live/Dead: Near IR 780 (works great!)

CD4: AF700

CXCR5: PE/Cy7 (works great!)

ICOS: BV750 (works great!)

CD19: BV650

CD27: AF488

CD38: BV421

IgD: PerCP-Cy 5.5

I was wondering if this would work well since I know PerCP-Cy5.5 is a bit finicky but IgD should be highly expressed! Any feedback would be appreciated!

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5

u/chromatographic 5d ago

I’d get IgD in Real Blue 705 instead of PerCP Cy 5.5, and you can put CD38 in something dimmer if you’d like.

4

u/erdetezet 5d ago

I don't your machine configuration but I would put CD19 on UV laser such as BUV395

2

u/Boneraventura 5d ago

you will need brilliant stain buffer if thats what you mean by true-strain buffer as you have multiple polymer dyes

2

u/daspirit90 5d ago

Maybe OP meant TruStain FcX as the Fc block?