CITE-seq data visualization
1
0
Entering edit mode
14 months ago
chi.delta ▴ 40

Hello, I have produced quite some single cell CITE-seq data at the single cell level, using the 10x genomics platform and analyzed in Seurat. I am at the moment trying to visualize the protein markers from the CITEseq against to each other in a way that would resemble how we see and interpret FACS data (with that also one could possible "gate" on single-positive and double positive populations the way we do in FACS analysis). Many times when I plot the expression level(using the Seurat function FeatureScatter) after normalization what I see is a plot like this: enter image description here

In this case, it is hard to recognize the single positive and/or double-positive populations but when I apply a log10 transformation with the code below, I get a better picture for the double positives.

FeatureScatter(alldata,"NKG2D","LAG3",group.by = "exp",cols = "grey") + scale_x_log10(limits=c(0.2,5)) + scale_y_log10(limits=c(0.2,5))

enter image description here

However, after the log transformation, all the cells with expression value 0 for these markers would just go to -Inf (we can see them stuck on the axes). Is there a way to correct for this?

For the same reason (generation of -Inf values), I also get this error after applying a geom_density_2d to the plot:

Caused by error in `MASS::kde2d()`:
! only finite values are allowed in 'lims' 
8: In min(x, na.rm = na.rm) :
  no non-missing arguments to min; returning Inf
9: In max(x, na.rm = na.rm) :
  no non-missing arguments to max; returning -Inf
10: In max(f) : no non-missing arguments to max; returning -Inf

Is there a better way to show these data? Is there a way to go for a bi-exponential correction as in FACS?

immunology scRNAseq FACS 10x CITE-seq • 536 views
ADD COMMENT
0
Entering edit mode
14 months ago
zau saa ▴ 150

Using sqrt transformation can avoid the problem and may produce similar graph.

ADD COMMENT

Login before adding your answer.

Traffic: 1822 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6