mass channel spillover
I was facing a problem in my last CyTOF run and was hoping someone could have ideas on that. I ran the same healthy blood donor PBMCs in 2 experiments 4 days apart, the cells were prepared the same way. I used the same antibody mastermix for both experiments, meaning that the mastermix was stored for 4 days in the fridge (diluted in Maxpar cell staining buffer). The samples were acquired on a CyTOF2 at a similar acquisition speed of about 400-500 events/sec. In the second run I see spillover from the 141Pr channel to some other channels it should not interact with (see attachment; no spillover to channels other than the plotted ones). Could this be due to instability of the antibodies when storing the mastermix? Does anyone have experience on storing pre-pipetted antibody mastermixes for several days?
There was an instrument wash between both runs leading to a higher Tb mean dual count, but this does not explain what I am seeing.
Any input will be welcome, thanks a lot in advance!
-Astrid