Fri May 06, 2016 6:57 pm by mkunicki
Hi Nick,
Funny bumping into you on Cytoforum!
I was wondering if you ever came to a conclusion on that CD127hi double positive population?
I have seen the same sort of staining (as shown here), but I see it with only one of my frozen healthy donor samples. The donor had other irregularities such as low CD4 count, and high %CD161+ in CD8 T cells which left me to believe this donor may have been ill at the time of blood draw. Considering isotopic impurity, doublet-discrimination, recurrence between experiments, and running multiple healthy donors in parallel, I believe this CD127+++ population had something to do specifically with this donor or sample prep.
I am curious if you'd be willing to discuss your sample processing & freeze/thaw protocol? It would be interesting for our group too to pin-point this issue. For example, one thing we see is perturbations in CXR or CD127 signal with longer/stronger fixation steps, and freezing has had a similar affect (al beit to a lesser extent).
Hope this helps,
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- CD127+++ events
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- Doublet-discrimination
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