Correlative Optical Tweezers-Fluorescence Microscope from LUMICKS for Basic and Translational Research at Georgetown University - SUMMARY This proposal is for the acquisition of a high-end optical tweezers with single fluorophore detection to be placed in a shared facility at Georgetown University, and to be used by researchers at the College of Arts and Sciences and the Georgetown University Medical Center. The proposal contains projects from 10 investigators, funded by NIH, NSF or private institutions, and is requesting a Lumicks C-Trap Dymo 300, an ultra-high resolution dual- trap optical tweezers coupled to fully correlated single-molecule confocal fluorescence detection of three colors and advanced integrated microfluidics. This is the only commercially available system with correlative force- distance measurements including the detection of single fluorophore emission or FRET, and thereby representing the state-of-the-art instrumentation in single molecule biology and biophysics. At present no such capability exists anywhere in Washington DC; the closest systems (NIH in Bethesda, Johns Hopkins University, Baltimore) are inaccessible or lack the essential 561 nm laser required by several of our researchers. This leaves our scientists either queuing for limited time slots in Baltimore or reformulating experiments around technology we have, often at the cost of novelty and impact. Installing a C-Trap Dymo 300 on campus therefore closes a critical regional gap and positions Georgetown as the natural hub for correlative force-fluorescence studies in the greater Washington DC area. Currently, Georgetown University has a single, home-built optical tweezers (without fluorescence detection or microfluidics) in a research laboratory in the Departments of Chemistry. The in-house expertise in single molecule manipulation methods positions us well to take full advantage of correlative force-distance and fluorescence measurements, a combined functionality that allows asking and answering new questions of ongoing and funded projects, but which is not currently available at Georgetown University or indeed within the capital district. Several projects will directly benefit from the C-Trap Dymo 300 and the specific configurations requested, including the development of fluorescence-based biosensors to visualize and quantify RNA dynamics in living cells (Braselmann), examining RNA binding to small molecules (Nair), studying IDPs and condensates (Nair, Metallo, Levi), characterizing organelle interactions (Levi), dissecting factors that govern tissue development, dysfunction and repair (Alimperti), quantifying dynamics, energetics and mechanics of protein kinase activation and regulation (Maillard), uncovering homeostatic mechanisms that stabilize neural circuits (Wang), among others. Altogether, the C-Trap will advance the research projects outlined here, and bridge new basic and translational investigations between the college and medical school at Georgetown University. This addition will make the facility the only one in the Washington, D.C. equipped with C-Trap instruments, positioning Georgetown University as a regional leader in single-molecule research.
