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slowkow avatar slowkow commented on May 27, 2024

Hi Kathrin, thanks for the question.

It sounds like you are giving the PCA coordinates of cells as input to Harmony. Is that right? If so, please see the documentation for your PCA function to get the variance explained for your PCs.

For example, you might be using the PCA function from the sklearn Python package:

https://scikit-learn.org/stable/modules/generated/sklearn.decomposition.PCA.html

Here is a detailed tutorial that shows how to use that function and find the variance explained:

https://jakevdp.github.io/PythonDataScienceHandbook/05.09-principal-component-analysis.html

In the issue you linked to, Ilya mentions that he added standard deviations of Harmony output to the Seurat V3 object. Here's the code for that:

https://github.com/immunogenomics/harmony/blob/88b1e2a1e95306cd4531eaf89ad98712af05ca99/R/RunHarmony.R#L149-L154

    harmonydata <- Seurat::CreateDimReducObject(
      embeddings = harmonyEmbed,
      stdev = as.numeric(apply(harmonyEmbed, 2, stats::sd)),
      assay = assay.use,
      key = reduction.save
    )

You can see that it's applying the sd() function to each column of the Harmony output matrix.

Maybe that's what you're looking for?

In Python, you can use numpy to get the standard deviation of each column of a matrix:

https://numpy.org/doc/1.18/reference/generated/numpy.std.html

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kathrinjansen avatar kathrinjansen commented on May 27, 2024

Thanks for the quick and detailed answer! I will calculate it using some of your suggestions.
May I ask your opinion on something related to this? Would you usually select the number of PCAs to use before running harmony (e.g. do an elbow plot) or is it acceptable to do a component selection post running harmony?

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slowkow avatar slowkow commented on May 27, 2024

I like to run multiple analyses with different parameters to get some sense of how they can affect the final results. The methods section of published papers can occasionally be helpful to see what strategies people use to choose parameters.

For general questions, you might consider asking for analysis advice on Biostars.

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kathrinjansen avatar kathrinjansen commented on May 27, 2024

Thanks for the answer, sorry if I hadn't phrase the question very clearly: it was specifically about harmony component selection. No worries though, you've answered my original question. I'll close this now, thanks again for the great tool & your help!

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