Hello, everyone!
I'm a new user of the Cytek Aurora and appreciate any guidance you can offer. I have some precious patient samples that I need to analyze to detect intracellular proteins while also separating different cell populations. My separation of various cell types from these samples works really well with beads as single-stain controls. However, I have a limited number of cells available (2 million per patient) and need to set up FMO (Fluorescence Minus One) controls.
Given that I can’t acquire more cells from these patients, would it be acceptable to use normal PBMCs (Peripheral Blood Mononuclear Cells) as physiological controls to set up my FMO gates?
Thank you in advance for your help!
Yes, normal PBMCs are definitely suitable for setting up your FMO control and for your single stain set up for compensation, but you also need to make sure that all markers can be stained by your normal PBMCs even if their staining intensity is not the same as patient PBMCs (at least positive). PBMCs should also be used for your single stains because you need to collect the background autofluorescence in all peak channels on Cytek Aurora, which cannot be replaced by beads as they have very distinct autofluorescence profile across all peak channels.
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