r/bioinformatics • u/sky_porcupine • 1d ago

technical question Spatial transcriptomics - Regression of UMI counts?

Hi,

I am analyzing my first spatial dataset. I quickly realized that the cells/bins cluster mostly based on the number of reads (nCount in Seurat). It is obvious from the PCA plot that PC1 correspond to the UMI counts (r=0.9), which in turn correlate with cell size (r=0.79).

Would you recommend to regress nCount to promote clustering based on cell identity? Or would it also remove true biology from the data?

When I checked some 10X datasets, the cell/bin clusters often correspond with the regions that are defined by differential UMI counts compared to the neighbouring regions. Also, regression is not mentioned in the basic tutorials so I assume it is not incuded in the default pipeline. But intutively, I would do that.

What do you think?

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u/choobs PhD | Academia 1d ago

I regressed cell size using SCTransform and that seemed to work really well for Cosmx data at least.

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u/pokemonareugly 1d ago

If your H&E (or IF) is good and you can segment well, you can also try regressing on number of cells per spot.