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

这个奖项是为了表彰生物发光比成像显微镜（BRIM）的发展。BRIM将同时记录和成像具有荧光素酶和光蛋白等生物发光报告的微观标本中两种不同波长的单个发光光子。BRIM遵循研究人员先前开发的生物发光共振能量转移（BRET）技术。然而，BRET发光传感器缺乏低光单光子灵敏度的光谱分辨显微成像。基因编码的生物发光报告构成了强大而灵敏的传感器的基础，可以记录基因表达、动态蛋白-蛋白相互作用和细胞内钙浓度的瞬时变化。这些生物发光报告者将被BRIM检测和成像。该开发项目解决了对微创生物传感器的需求，该传感器可以实时记录活细胞中的信号转导事件。BRIM系统的原型将提供给其他机构的合作研究人员。该奖项将为生物医学工程师和分子细胞生物学家组成的跨学科团队中的研究生和本科生研究实习生提供研究培训。

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.