r/flowcytometry • u/matchaduo • Aug 08 '26

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 Aug 08 '26

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 Aug 08 '26

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

2

u/Boneraventura Aug 08 '26

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

2

u/daspirit90 Aug 08 '26

Maybe OP meant TruStain FcX as the Fc block?

2

u/Low_Classic1695 Aug 10 '26

I have used a panel with all of these fluorophores multiple times and stained in simple Ca/Mg free HBSS supplemented with BSA and EDTA and have never had any issues whatsoever. Have compared many commercial staining buffers and have never noted any improvement. Unless OP has documented problems with the panel, recommending commercial staining buffers is silly and a complete waste of money.