QuestionsCategory: Single-Cell RNA-SeqWhy don’t the scRNA transcript counts match up with the results I found with flow cytometry?
wongnw@nih.gov asked 3 months ago
1 Answers
kellymc@nih.gov answered 3 months ago

The most likely difference between an scRNA-Seq data and flow cytometry data (assuming) is because there may be a difference in the quantity of transcript for a gene and the amount of protein being measured. Poor correlation is not always unexpected – they are different parts of gene expression stages and so difference could be due to a time delay between level of transcripts and level of protein present. You may also not need a large number of transcripts to make a lot of protein, and the half-life of transcripts and proteins may be different.  
There are also potential technical reasons for a why the RNA may not match protein levels. You may have ‘drop-outs’ in detection of the RNA because the reverse transcription was not efficient and/or because we are at the lower detection limit. Detection sensitivity of the protein as a more direct measure may be higher on flow.