Cytometry and Antibody Technology

Somewhat of a mea culpa

by | Apr 1, 2024 | ChUG Podcast, Learn: Spectral Cytometry | 0 comments

The latest ChUG podcast is out where we discuss a number of topics. But I wanted to revisit the unmixing with beads issue that we discussed in a previous post. So it turns out beads create unmixing matrix that do not correct spectral artifacts completely. I’m guessing researchers using flow cytometry have been aware of this for some time and have simply adapted to the problem by adding compensation on top of the unmixing matrix in order to smooth out the data presentation. On the podcast, Ryan explains his point of view on the matter, and I think it is shared by a lot of people in the field. Meanwhile, flow core facilities such as mine have pretty much trumpeted best practices as the solution to that problem: if your uminxing is not working for you, it’s because your controls are not good enough. This can be true in many cases, but it does blinds us to the fact that at some point, controls are going to be what they are and we still need to move forward. In other words, if your controls are as good as you can possibly make them, how do you correct the unmixing issues that you’re still experiencing? I think core facilities, flow cytometry training programs, and manufacturers have not provided a clear answer at this point, and that should change. What is the teachable skill that we can share to help our users? There a few ideas worth exploring in our podcast, and I’m sure more will emerge at some point.

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