Refeyn TwoMP Mass Photometer with MassFluidix Microfluidic High Concentration (HC) Technology - PROJECT SUMMARY Our understanding of fundamental biological processes is both driven and limited by our ability to visualize macromolecular structure and organization. Access to rapid characterization of proteins and nucleic acids at the single-molecule level is now essential for scientists to push the frontiers of biological research and is critical for the application of this knowledge for the improvement of human health. This proposal requests funds to purchase the TwoMP Mass Photometer with MassFluidix Microfluidic Technology (Refeyn, Inc.) for the analysis of biological molecules in the size range from kilodalton to megadalton. The fast imaging and high-resolution capabilities of the TwoMP are also designed to overcome the speed and sample abundance limitations of single-molecule imaging. We have identified the TwoMP as the sole commercial product which meets our requirements, is state-of-the-art and cost-effective. This instrument will be a shared resource in a well-established core facility at Stanford, the Macromolecular Structure Knowledge Center (MSKC). The TwoMP mass photometer will support more than twenty cutting edge health care NIH projects described in this proposal. These projects cover a wide range of topics, including: energy metabolism (Long); drug discovery (Gray); cholesterol (Welander); gene therapy (Qi); insulin-independent pathways (Svensson); neurotransmission (Maduke); biomaterials (Mai), brain development (Fame); signaling (Jackson); peptide biomimicry (Barron); receptors (Barnes); RNA (Das); transcription factors (Khavari); oxidative damage (Wang); allergy (Jardetzky); mRNA (Martinez); genome assembly (Straight); autoantibodies (Lanz); nanoparticles (Sharaf); chromatin architecture (Altemose); celiac disease (Khosla); pandemic viruses (Lin); and vaccine development (Kim). These studies address critical functional and structural questions about fundamental biological processes and span NIH research areas with implications for diverse aspects of human health and disease, including diabetes, cancer, dementia, obesity, Alzheimer's disease, influenza, and pandemic viruses. The speed and low sample requirements of the TwoMP instrument are critical for all these projects, where sample scarcity and instability are often a significant challenge. These combined capabilities will drastically enhance our NIH funded faculty ability to conduct biophysical research by providing a shared resource that meets their biomolecular analysis needs.