High-Throughput Screening (HTS)

Automated robotic experimentation testing tens of thousands to millions of chemical or genetic perturbations against a biological target.

Specifications & Metrics

type: Hit Discovery & Automation
readout: Fluorescence intensity, FP, FRET/TR-FRET, luminescence, or high-content automated microscopy
measures: Primary hit activity (% inhibition or activation) and quantitative concentration-response (qHTS) profiles
format: 384-, 1536-, or 3456-well microplates and droplet microfluidics integrated with liquid-handling robotics
pioneer: Pharmaceutical screening facilities in the late 1980s–1990s; quantitative HTS (qHTS) established at NIH NCATS

Robotic HTS platforms copy nanoliter-to-microliter volumes from compound stock libraries into 384- or 1536-well assay plates, testing up to 100,000 compounds per day (with ultra-high-throughput screening, uHTS, exceeding 100,000 per day).
Primary screening hits are cherry-picked from stock wells for secondary confirmation, counter-screening against assay artifacts, and multi-point IC50/EC50 titration to establish initial structure–activity relationships (SAR).
Quantitative high-throughput screening (qHTS), pioneered at the NIH National Center for Advancing Translational Sciences (NCATS) in 1536-well plates, tests every library compound across four to five orders of magnitude of concentration in the primary run.