During this NIH SBIR Phase II project, Dakota Technologies, Inc. (DTI) will complete the development of a revolutionary laser-induced fluorescence instrument that provides the flexibility and convenience of research-grade microplate readers, but collects data at speeds typical of high throughput screening. The Phase I project investigated and established proof of concept for novel fluorescence lifetime measurements applied to microplate reading. DTI has the ability to measure very high quality fluorescence decay curves two orders of magnitude faster than the competition. The Phase I research also demonstrated the value of globally analyzing fluorescence polarization data for improved characterization of drug-protein binding and an extremely sensitive means to monitor DNA hybridization without the need for labeling either probes or targets. By the end of Phase II, DTI will be able to acquire a complete fluorescence decay curve (waveform) every time a high repetition rate (10,000 pulses per second) solid-state microlaser fires. The compact passively Q-switched microlaser provides high peak power, short pulse duration, and outstanding shot-to-shot stability. As a consequence, the dominant influence on data quality is photon statistics, not noise of the measurement system. Compared to TC-SPC, where data is acquired for one fluorescence photon per 100 laser shots or more, we record the response for tens to hundreds of fluorescence photons per laser shot. Important Phase II activities include transforming the Phase I proof of concept microplate reader into a robust engineering prototype and developing optimized assays that take advantage of the system's unique features.
