Development of a Bioluminescence Ratio Imaging Microscope (BRIM) to Visualize Biosensors for Signal Transduction Events in Living Cells in Real Time

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This award is for the development of a Bioluminescence Ratio Imaging Microscope (BRIM). The BRIM will simultaneously record and image single luminescent photons at two different wavelengths in microscopic specimens that have bioluminescent reporters such as luciferases and aequorin photoproteins. BRIM follows the investigators' previous development of the Bioluminescence Resonance Energy Transfer (BRET) technique. However, the BRET luminescence sensor lacks spectrally resolved microscopic imaging with low-light single-photon sensitivity. Genetically encoded bioluminescent reporters form the basis for powerful and sensitive sensors to record gene expression, dynamic protein-protein interactions, and transient changes in intracellular calcium concentration. These bioluminescent reporters will be detected and imaged with the BRIM. This development project addresses the need for minimally invasive biosensors that can record signal transduction events in living cells in real time. The prototype BRIM system will be available to collaborating researchers at other institutions. The award will provide research training for a graduate student and for undergraduate research interns in an interdisciplinary team of biomedical engineers and molecular cell biologists.