The new Analyst™ GT is a multi-mode plate reader supporting all five leading technologies: fluorescence polarization (FP), fluorescence intensity, time-resolved fluorescence (TRF), absorbance and luminescence. It can be easily integrated into both workstation and robotic environments for HTS for all major non-radiometric applications. Analyst™ GT is based on second-generation SmartOptics™, a patented system that combines light sources, optical components and detectors together for fast high-precision measurements in 96-, 384-, and 1536-well microplates. Applications include kinase and photophase assays, receptor ligand binding assays, protease assays, cAMP, and cGMP, signal transduction, reporter gene assays and many others. In this study, we not only optimized measurement parameters on Analyst™ GT for all five modalities but also compared instrument settings and performance with the first generation Analyst HT and Acquest.