SBIR Phase I: CryoPREP-EM: Time-Resolved Sample Preparation for Single-Particle Cryo-Electron Microscopy (Cryo-EM)

This Small Business Innovation Research Phase I project will evaluate the technical feasibility and commercial viability of a novel sample preparation method for cryo-EM (cryogenic electron microscopy) studies. As a rapidly growing technique in the structural biology field, the cryo-EM market as a whole is estimated to be more than $5 billion a year, with > $500 million attributed to the sample preparation space. In spite of an overall revenue growth rate of more than 70% per year, cryo-EM sample preparation remains an area that has yet to see any major technological breakthroughs commercialized in the past decade. In fact, the current industry solution is reported to have a failure rate of > 95%. By providing a sample preparation system that actually works, this Small Business Innovation Research (SBIR) Phase I project will significantly reshape the massive and rapidly expanding cryo-EM market and allow scientists to more readily collect detailed protein structures in their native state. This new knowledge on the molecular basis of cells and of life itself will be used to facilitate as well as accelerate the development of new pharmaceuticals as well as therapeutic treatments for diseases and thus greatly benefit human health in the long run. The intellectual merit of this project is the development of a cryo-EM sample preparation device built on the application of state-of-the-art microfluidic jet systems to deposit and vitrify protein solution on EM grids in milliseconds, which is at least 1000x faster than the current methods. Furthermore, it offers scientists the novel ability to control the thickness of the vitrified aqueous layer and, overall, more control over the sample vitrification process. With this new sample preparation device, a larger region of the cryo-EM grids will have intact protein molecules with the right layer thickness for structural determination, allowing researchers to save time and money in terms of both grid preparation and screening in a high-throughput manner. Currently, the proposed technology has not been tested nor used anywhere. Therefore, the first and foremost goal of this SBIR Phase I project is to better understand and well characterize the technical capabilities and limitations of this proposed innovation, with additional R&D work needed before product commercialization, which is expected in five years. The project will focus on defining and optimizing both the geometric and experimental setup in order to de-risk the technology. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.