Thu Jul 23, 2020 4:41 pm by mleipold
Hi Fanta,
Thanks for the panels.
I would strongly recommend that you put the metals in numerical order when you are looking for spillovers. Spillovers are primarily due to elemental impurities of the same mass, isotopic impurities of the same element, and oxide spills. Having them in order makes it a lot easier to see if you have the same element, calculate M+16, and other things.
I have rearranged your table in order, where the green are the 3 new markers and the pink are the two channels in your bivariate plots.
I am assuming that you watched the sample being acquired, so that you can rule out the possibility that the increased background is caused by streakiness due to debris or other sample running or sample quality issues.
Assuming that's true, looking at your panel and your bivariate plots, my guess is that you're getting isotopic impurity spill from your new TIM4-154Sm marker back into the CD3e-152Sm channel. They're the same element, and the next nearest mass, so they would be the most likely to spill.
The simplest way to check this is doing a MMO experiment, where you leave out the TIM4-154Sm but keep all the other 28 antibodies.
Mike