We compare virtual labelling performance across eight alternate molecular marker channels (8-plex, including: α-Tubulin; DNA (DAPI); CoxIV; fibrillarin; GM130; F-actin (phalloidin); β-Catenin; NF-κB-p65) concurrently labelled via the iterative indirect IF imaging (4i) experimental multiplexing protocol33 in 45,780 DU145 prostate cancer cells (Fig. 2B) and use differential interference contrast (DIC) images as the label-free component. The gene discussed is NFKB1; the disease is prostate cancer.