broadinstitute/CellBender

Problems interpreting results

MarP2606 opened this issue · 0 comments

Hello,
thank you for the tool!
I am a first time user and I am struggling a bit to interpret the results plots and to change the settings adequatly for the next run.

We performed songle-nuclei RNA sequencing from tissue. Therefore we expect high ambient RNA levels.
The cellranger pre-processing recovered about 9.800 cells which fits with our expectations of targeting 10.000 cells.
cellranger_report

I ran cellbender with the default settings.
I think the learning curve looks ok from what I've gathered from other issues.
learning_curve

I don't know what to expect for the top genes removed, but could it be that the fractions removed seem a bit low?
genes_removed

Further, cellbender identified 5 times more cells than expected.
cell_probability

So now I am a bit unsure which settings are the appropriate ones to change.

  1. Should I change the FPR? Would 0.05 be the next try?
  2. Should I change total-droplets-included (higher?) and expected-cells (10.000)? I also saw that changing the parameter low-count-threshold was suggested, which would be probably 1.000 in my case. What of those two options is better?

Thank you for your help!