high-quality data at the HTS level is vital to the success of the subsequent steps, which are much more expensive and lower in throughput. The HTS data must be of sufficiently high quality, with low incidence of false positives and negatives, in order to establish an effective roadmap for the subsequent processes. Too often, the emphasis is on rapidly screening huge libraries (''quick and dirty''), producing inefficiencies in subsequent steps. By taking advantage of new fluorescence lifetime measurement technology, powerful new assays are enabled. Our premise is that time-resolved FRET (TR-FRET) is so highly advantageous over intensity-based assays that it should always be the first choice. TR-FRET on the nanosecond time scale is not the norm now. The most familiar form is live cell assays with genetically encoded fluorescent proteins, which are generally approached by FLIM. However, the low speed of FLIM makes it applicability for HTS very limited. Our solution is to employ a ''Cells-in-Wells'' strategy in which the collective response of hundreds of cells are monitored simultaneously in an non-imaging format. We will present data that demonstrates the conversion of assays that are useless for HTS in an intensity mode (z' < 0) to very robust assays (z' > 0.7) in fluorescence lifetime mode.
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