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SamGG avatar SamGG commented on September 17, 2024

Hi,
IIUC, you are trying to map a set of 8 FCS to an already computed FlowSOM result. To do so, the 8 FCS must be mapped to the FlowSOM result. Read the following and the overall step-by-step pipeline which is awesome.
Best.
https://github.com/saeyslab/FlowSOM_protocol/blob/d2fbbfe595baf6996be8966b3f7279f7413151fe/R/FlowSOM_protocol.R#L460-L471

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maiabennett avatar maiabennett commented on September 17, 2024

Hi,

To clarify, the FlowSOM object I am using is constructed of a concatenated .FCS file of the 8 individual files. I am attempting to run an analysis following part 9, 'Comparing different groups', of the FlowSOM vignette. I do not want to create a new FlowSOM result, but to compare the differential percentages of clusters present between two treatment groups. I have read through the code you have provided as well as the R package documentation and do not believe the highlighted NewData function is applicable, as it does not return the necessary percentages matrix. Do you have any further suggestions? I do agree that your pipeline is very good.

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SamGG avatar SamGG commented on September 17, 2024

This pipeline is from @KatrienQ, one of the developer of FlowSOM, not me. The dev will help you better than me.

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SofieVG avatar SofieVG commented on September 17, 2024

Can you just confirm what the column names from your original training data were (e.g. by colnames(flowSOM.untreated.res)) and what the column names from your new data are (e.g. colnames(untreatedset[[1]]) )?
I am thinking maybe you aggregated files in your baseline.untreated and might have some additional columns there, not present in your untreatedset ? If it is a concatenated file with e.g. a SampleID or File column, you might want to specify the colsToUse parameter in FlowSOM to make sure those columns are not used for clustering.

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maiabennett avatar maiabennett commented on September 17, 2024

@SofieVG, thank you! I was able to trace back the error source. It was due to differences in parameters between the concatenated file and its component individual files. Thanks again for your time!

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