Chatomics Field GuideWhat They Don't Teach You

Topic

Pseudoreplication

Are my cells (or spots, or technical replicates) really independent samples?

Treating 5,000 cells from 2 donors as 5,000 replicates produces p-values of 1e-200 for effects that would not replicate in a third donor. The unit of replication is the biological sample; pseudobulk aggregation or mixed models restore it.