SBIR Phase II: Novel platform for the preparation of samples for cryo-TEM imaging: capturing biomacromolecules repeatably in vitreous ice without artifacts

The broader impact of this Small Business Innovation Research (SBIR) Phase II project is a new way to prepare samples for life sciences research by addressing the major bottleneck in the associated workflow. The proposed work advances a new method to prepare biological samples for analysis to develop crisp, high-resolution 3D models of molecules. This will result faster acquisition of higher quality data and better 3D models of proteins and other cellular components, providing new insight into human biology and improvements to human health. The proposed project aims to solve the largest issues currently experienced in cryogenic Transmission Electron Microscopy (cryo-EM) sample preparation. Current sample preparation technologies are unreliable and require preparation of excess grids to acquire enough quality data for a 3D model of a biomacromolecule. Many grids are of poor quality and generate insufficient data for a high-resolution protein model; or samples may contain artifacts from interaction at the air/water interface, reducing the ultimate resolution. This Phase II project will advance an innovative design concept to repeatably prepare high quality samples with a blotless sample preparation technique. The proposed work will explore the trade space and optimize process parameters for image resolution, leading to an advanced prototype. 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.