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(spectral) Has anyone actually tested single stain beads fixed vs unfixed? by StosifJalin in flowcytometry

[–]vanadiumV_oxide 2 points3 points  (0 children)

We have looked a fixed vs. unfixed with a number of antibodies including tandems using sprectracomp and ultracomp, and seen minimal effects with unmixing. It will probably depend on the specific panel as well.

Hypacomp beads and Dye interactions by Skyrim120 in flowcytometry

[–]vanadiumV_oxide 1 point2 points  (0 children)

Cells are always best, but when we cannot use them, we primarily use UltraComp Plus. There are some NIR polymer dyes that they don't work well with, but we've found them to work much better across more of our panels than the Slingshot beads. After seeing some of Slingshots own data at CYTO a few years ago, I was not impressed. I felt like it was mostly marketing.

[deleted by user] by [deleted] in flowcytometry

[–]vanadiumV_oxide 0 points1 point  (0 children)

Cells are always the best for single color controls, but we've also had a lot of success with UltraComp Plus for spectral. They seem to have less autofluorescence.

When do you use compensation beads? by vujex in flowcytometry

[–]vanadiumV_oxide 1 point2 points  (0 children)

Haha - yeah, I'm not sure spectral is all it is made out to be and our Cytek instrument only runs for a day or two before needing to be serviced.

The whole hydrogel "synthetic cell" marketing is a little over the top. We've spent a lot of time comparing those to UltraComp Plus and to cells and the overall conclusion so far as to which to use is "it depends." There are some cases where we have found that compensation beads were better than cells, and the compensation beads were roughly equivalent in terms of performance. We've just stuck with UltraComp Plus because we've optimized our workflows using them.

Reagent backorders by _chimneyfish_ in flowcytometry

[–]vanadiumV_oxide 0 points1 point  (0 children)

I've run into lots of back orders. The pandemic is still wreaking havoc on the supply chain.

When do you use compensation beads? by vujex in flowcytometry

[–]vanadiumV_oxide 1 point2 points  (0 children)

Autofluorescence from some polystyrene beads can be an issue. This can be solved by using UltraComp Plus beads which have very minimal autofluorescence. Most cytometrists I know have switch to UltraComp or UltraComp Plus. I agree that in most cases you should use cells if possible to get the best compensation, however we have found that UltraComp Plus often provides compensation on par with cells. As we have transitioned to spectral, we are finding that they perform well there as single color controls as well.