Thu Feb 05, 2015 4:40 pm by AdeebR
Hi Joanne,
I've done some testing of the SmartTube buffer for preserving whole blood (without the base station).
It certainly seems to do a nice job of preserving the cells, and intracellular targets. Recovered frequencies of major subsets were similar before and after cryopreservation. However, the buffer seemed to degrade a number of surface antigens that are of interest to us (essentially all chemokine receptors) so we ultimately decided that it wouldn't be that useful for the study we were considering it for.
I do a lot of myeloid work with whole blood. Here's my "core" myeloid backbone for whole blood:
CD66b-152Sm
CD16-148Nd (gives strong signal on neutrophils, but never stains other subsets as well in whole blood as in PBMCs)
CD14-160Gd
CD1c-153Eu
CD141-144Nd
CD123-151Eu
HLA-DR-174Yb
CD11c-159Tb (not critical since it's expressed fairly promiscuously by human monocytes/DCs)
CCR3-164Dy (also not critical, but useful for definitively identifying eos/basophils - needs to be stained prior to fixation).
I also typically include CD19, CD3, and CD56, which are useful as exclusion parameters when identifying DCs.
Also note that whole blood is not ideal for identifying DCs on the CyTOF because you need to collect a lot of events to compensate for all the granulocytes.
Adeeb Rahman
Icahn School of Medicine at Mount Sinai, NYC