Readily deployable technology for reproducible cryogenic sample time-resolved electron microscopy (cryoTREM)

SUMMARY This advanced technology grant focuses on enhancing cryogenic sample time-resolved electron microscopy (cryoTREM) through the development of a patterned LED/LCD-based illumination system capable of capturing multiple temporal states on a single EM grid with 4 orders of magnitude time range from 1 to 10,000 milliseconds. This system will be developed and deployed in producing time-resolved data sets for several different biological systems with intermediate states with varying degree of conformational magnitude. The resultant data will be integrated with a variety of data processing methods to determine ensembles of time-resolved structures, which can be leveraged to understand molecular mechanisms. These time-resolved EM experiments will be conducted on multiple biological systems and processes including ligand-gated ion channels, a lipid transferase, the V- ATPase, and synaptic vesicle fusion.