Overview of hcHCR. (A) Human primary cells are purified and plated in 384-well imaging plates, followed by in vitro treatment, RNA HCR, and high-content quantitative imaging at single-cell resolution. (B) Representative high-content image analysis of hcHCR data: segmented nuclei regions, cell body regions, and HCR spot borders are shown in pseudocolors. Magnification: 60×. Scale bar for full FOV: 20 microns. Scale bar for inset: 10 microns. (C) Analysis and visualization of hcHCR data. (Left) A representative single-cell density plot, comparing the distribution of HCR spots per cell between two conditions. (Center) A representative display of the mean and SEM for HCR spots per cell over time. (Right) High-throughput HCR data can be analyzed and gated with standard flow cytometry software. A representative histogram of cell counts by HCR spots per cell, with two gates, is shown.
