Abstract:
The past decade of advances in molecular biology has revealed that the cell comprises a complex system of networks on the scale of atoms, molecules, and organelles. The next breakthroughs in life science research, in academic labs and as applied to drug development, and other translational disciplines, will depend on the ability of physicists and engineers to unravel the complex biophysical phenomena that underlie cellular function with greater resolution. Most currently available technologies rely on ensemble population measurements which frustrate the absolute quantitation which is required to reveal the true complexity of life at the molecular scale. In this talk, I will introduce Convex Lens-induced Confinement (CLiC) microscopy, a general method to image molecular interactions one molecule at a time, while emulating 'cell like'conditions, with precision and control. Because it mechanically confines molecules in the field of view, CLiC eliminates the need to tether molecules thereby avoiding the complexity and potential biases. By visualizing the individual trajectories of many molecules at once, and for long time periods, CLiC allows us to investigate important biophysical questions about molecular behaviour, such as how higher-order DNA structures can regulate the dynamic unwinding of specific target sites within a crowded environment, and the kinetics of binding to these sites.Beyond discussing new insights from CLiC into the statistical mechanics of DNA, I will discuss key applications of our work in drug development, including visualizing protein aggregation, nanoparticle dynamics, and CRISPR-Cas9 targeting dynamics.