Fluorescence resonance energy transfer (FRET) techniques offer a new and unique approach for evaluating molecular interactions within cells, and have led to breakthroughs in our understanding of DNA structures. The objective of the proposed research is to extend the current FRET techniques to offer the capability for continuous observation at a high temporal resolution. The innovative aspect of the method described in this proposal is that it will minimize the probability of irreversible destruction of the donor or acceptor, which cannot be achieved with the current photobleaching FRET techniques.
The proposed concept will be implemented on a confocal microscope imaging system allowing the mapping of energy transfer efficiencies on a pixel-by-pixel basis. The method will be used to study nucleic acid interactions, and protein-nucleic-acid interactions in an
interdisciplinary effort with other scientists in the host laboratory.