LUMICKS C-trap for Mechanistic Studies of Biological Systems at the University of Minnesota - Project summary This proposal seeks the acquisition of a LUMICKS C-Trap instrument at the University of Minnesota. The C-Trap is the only commercially available laser optical trap that combines TIRF fluorescence microscopy with integrated microfluidics in a user-friendly, reliable, engineered instrument. Single molecule force and fluorescence spectroscopies have transformed the fields of cellular and molecular biology, biochemistry, biophysics, and biomedical engineering, providing answers to crucial biological and health-related questions. These tools are currently limited to laboratories with specialized expertise and custom-built single molecule instruments. The C-Trap is designed as a turnkey system, enabling laboratories without specific single molecule force or fluorescence spectroscopy expertise to conduct advanced single molecule force, position, and fluorescence localization, as well as FRET analysis, with unparalleled spatial and temporal resolution. Key features of the C-Trap include dual optical traps for manipulating biomolecules with sub-pN force resolution/detection; 3-color laser TIRF and widefield fluorescence detection for visualizing biological processes; precise temperature control; micro and nano-stage control for exact sample positioning; laminar flow microfluidics for high sample throughput and varying ambient media while maintaining molecular interactions; and user-friendly software for integrated, controlled, and automated instrument operation. The intuitive instrument and software interface, along with the automation package, will enable non-expert users to run experiments and collect high-quality data with minimal training. The proposal represents five major users (all with NIH R01/R35 funding) who will primarily use the instrument, and eight minor users with NIH or other funding. These users come from four colleges across University of Minnesota (College of Biological Sciences, College of Science and Engineering, School of Dentistry, and the Medical School) and six different departments. The C-Trap will significantly advance the NIH-funded projects of these researchers by enabling high-impact single molecule mechanobiological and fluorescence studies on a range of important biomedical questions in areas such as cell surface mechanosensing, muscular dystrophy pathogenesis, cytoskeletal structure and dynamics, viral assembly and cell entry and nucleic acid processing. The major users and technical advisors possess extensive relevant expertise with the technology, and will provide guidance to all users. The instrument will be housed in the University Imaging Center, a Nikon Center of Excellence, which provides support and training for all of their instruments as well as expert advice and consultation, from experiment planning to image acquisition and analysis. Establishing this resource will significantly enhance numerous research programs at the University of Minnesota and contribute to the discovery of impactful scientific, biomedical, and healthcare-related knowledge.