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I'm working on a dataset where all cells have the expression of a given marker. I want to show this, per cluster, on a DotPlot; but leaving the default scale=TRUE argument gives me negative values in 2 out of 8 clusters. If I understand correctly, this means the gene has lower than average expression in these, when compared to the remaining clusters. Yet, I want to avoid the possible confusion for collaborators, reviewers, and ultimately, readers of the paper.
I'm not scaling the values (switching to FALSE). This is clear to me after some trials, I want the plot to be clear. Now, I've read the source code to get a grasp of what it means... I was afraid that turning this off would instead get me the geometric mean.
Since FetchData() has no layer argument defined, that's going to be @scale.data ... Right? Please confirm. And, looking to what DotPlot does, these are then potentiated (expm1), arithmetic mean is calculated, and log1p-transformed. Which is fine, but it took me a while to digest.
For the record, some grievances I had, were:
Shouldn't DotPlot call AverageExpression? If that were the case, I'd like to switch the layer there to have maximum control on what I'm plotting (e.g. via ellipsis, ...). By default, the function assumes data values need exponentiation (cfr. details at the reference manual)... anyway, averaging is not done by using this function and I found that to be confusing at first, so I wanted to complain :P bla bla DRY principles.
I'm using RNA assay, but the same observation applies to SCT, which is what I'm actually going to use across the whole pipeline for finding markers and identification of clusters' cell type. But for this plot, I had set off scaling and switched over to RNA. Else, I'd get zeroes (which, to my situation, is as bad as having the negative values).
Any advice will be appreciated. But mainly, I need to confirm the SeuratObj::FetchData behaviour. Thanks in advance!
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I'm working on a dataset where all cells have the expression of a given marker. I want to show this, per cluster, on a DotPlot; but leaving the default
scale=TRUE
argument gives me negative values in 2 out of 8 clusters. If I understand correctly, this means the gene has lower than average expression in these, when compared to the remaining clusters. Yet, I want to avoid the possible confusion for collaborators, reviewers, and ultimately, readers of the paper.I'm not scaling the values (switching to
FALSE
). This is clear to me after some trials, I want the plot to be clear. Now, I've read the source code to get a grasp of what it means... I was afraid that turning this off would instead get me the geometric mean.Since
FetchData()
has nolayer
argument defined, that's going to be@scale.data
... Right? Please confirm. And, looking to what DotPlot does, these are then potentiated (expm1
), arithmetic mean is calculated, andlog1p
-transformed. Which is fine, but it took me a while to digest.For the record, some grievances I had, were:
Shouldn't DotPlot call
AverageExpression
? If that were the case, I'd like to switch thelayer
there to have maximum control on what I'm plotting (e.g. via ellipsis,...
). By default, the function assumes data values need exponentiation (cfr. details at the reference manual)... anyway, averaging is not done by using this function and I found that to be confusing at first, so I wanted to complain :P bla bla DRY principles.I'm using RNA assay, but the same observation applies to SCT, which is what I'm actually going to use across the whole pipeline for finding markers and identification of clusters' cell type. But for this plot, I had set off scaling and switched over to RNA. Else, I'd get zeroes (which, to my situation, is as bad as having the negative values).
Any advice will be appreciated. But mainly, I need to confirm the SeuratObj::FetchData behaviour. Thanks in advance!
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