Lumicks C-Trap Single-Molecule System: Correlated Dual Optical Tweezers and Confocal Fluorescence Microscopy - Project Summary/Abstract This proposal seeks funding for a Lumicks C-Trap instrument for biomedical researchers at The Pennsylvania State University. The C-Trap, a unique commercially available optical trap instrument, integrates scanning confocal microscopy and microfluidics with a user-friendly interface. Correlated force and fluorescence spectroscopies have transformed single molecule studies in life sciences research, bridging high-resolution structures with their dynamical changes. Currently limited to specialized labs, the C-Trap is designed as a turn- key system, allowing non-expert laboratories to conduct cutting-edge single molecule studies with unparalleled spatial and temporal resolution. Key features include dual optical tweezers for piconewton force manipulation, three-color laser confocal fluorescence detection, precise temperature control, nano-stage positioning, laminar flow microfluidics for high sample throughput, a user-friendly Bluelake data collection and Pylake data analysis software. This proposal represents shared equipment use for ten major users and five minor users spanning five departments (Chemistry, Biochemistry and Molecular Biology, Bioengineering, Cellular and Molecular Physiology, Medicine, Microbiology, and Immunology), three colleges (Eberly College of Science, College of Engineering, and College of Medicine), and two campuses (University Park and Hershey). The C-Trap will advance fifteen ongoing interdisciplinary NIH-funded projects studying gene regulation, RNA and protein structure-function, mechanobiology, phase condensates, transcription, replication, translation, and DNA repair. The breadth of single molecules studies will encompass a diverse range, from molecular motors, cardiovascular proteins, cytoskeletal filaments, viral protein-RNA complexes, metalloproteins, LLPS in situ and nucleic acid processing enzymes, to multi-compartmentalized membraneless organelles and in vivo-like conditions. Scientific directors of the C-Trap core facility, Drs. Lee and Yennawar, will provide equipment oversight, expertise, and training to new users, while the Huck Institutes at Penn State will commit substantial institutional resources and support for effective utilization. The recent leasing of the C-Trap equipment has enhanced various research programs and centers at Penn State and synergized with other NIH S10 funded capabilities at the core, contributing to the discovery of impactful healthcare-related knowledge. Currently, there is no homemade optical tweezers setup in operation in any of the cores or any individual labs. The absence of commercially available instruments that match the unique capabilities of the Lumicks C-Trap across the twenty- four campuses of The Pennsylvania State University underscores the urgent need to acquire this advanced equipment. The C-Trap's high-resolution dual optical tweezers, ultra-stable force detection, five-channel flow cell microfluidics, and three-color imaging confocal microscopy system would significantly enhance the university's research capabilities, enabling cutting-edge studies that are currently unattainable with existing resources.
