High throughput screening using dynamic fluorescence

摘要

Multifrequency phase-modulation data for fluorescence lifetime and anisotropy decay standards have been obtained for liquid samples in 96-well microtiter plates. We coupled the bifurcated fiber optic probe of a fluorescence plate reader to a phase and modulation time-resolving fluorometer. Laser excitation was used and multifrequency data could be acquired quickly over a range of modulation frequencies from 15 to greater than 150 MHz, at each well position, through the use of multiharmonic Fourier (MHF) methods. We evaluated the system's performance for UV-excited fluorescence, with the goal of screening quality or properties of proteins or peptides by steady-state or dynamic intrinsic fluorescence intensity or anisotropy using the high throughput 96-well plate format.