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Multiplets after scrublet? #54

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rhlgyb opened this issue Jun 22, 2023 · 0 comments
Open

Multiplets after scrublet? #54

rhlgyb opened this issue Jun 22, 2023 · 0 comments

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@rhlgyb
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rhlgyb commented Jun 22, 2023

Dear managers of scrublet,

Thank you for your time. I will try to be concise.

The gist is, I am curious, with my limited knowledge, what other intepretations or hypotheses I could make other than these cells being multiplets...I'm sure I used scrublet correctly.

I have used

  • cellbender : to rid low quality cells filled with ambient RNA.
  • (afterwards) : cellbender filtered h5 file -> Seurat count matrix -> subsetted seurat object with scrublet barcodes
  • numerous combinations of normalization and batch correction methods
  • tried removing some blacklist genes before processing the data

However, I still end up with a number of cells from a specific sample that expresses the marker genes of fibroblasts, immune cells, epithelial and endothelial cells; pretty much all of the major cell types.
(cells were used for this analysis : percent.mt < 10~20, etc, 500 < nFeatures < 6000, 1000 < UMI)

Thank you so much for your help,

R.

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  • (above) colors represent respective samples

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@rhlgyb rhlgyb changed the title Cells in sample suspicious for multiplets after scrublet? Multiplets after scrublet? Jun 23, 2023
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