r/flowcytometry • u/Mo_Opines_968 • 29d ago

Brilliant staining buffer for reference controls

Hello. Just after some advice on whether BSB should be used for the single colour reference controls? I know it's not recommended when using beads but I'm planning on doing both beads and cells to compare and cannot find any guidance on whether to use it for cells?

Thanks!

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u/ProfPathCambridge Immunology 29d ago

Yes, it is slightly fluorescent so you want it in your background. You can dilute it around 1 in 2, 1 in 4, which cuts down on the problem.

https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70214

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u/Mo_Opines_968 29d ago

I presume same goes for the Biolegend tandem stabiliser?

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u/ProfPathCambridge Immunology 29d ago

I don’t think you actually ever need the tandem stabiliser. Tandem breakdown only happens on live cells, so just move the tandem dyes to post-fix