Flow Magnetic Tweezers: Imaging Rare Enzymatic Events with Single Molecule Precision

The application of forces and torques on the single molecule level has transformed our understanding dynamic properties biomolecules, but rare intermediates have remained difficult to characterize due limited throughput. Here, we describe a method that provides 100-fold improvement in throughput force spectroscopy measurements with topological control, which enables routine imaging 50,000 molecules 100 million reaction cycles parallel. This detection events life cycle cell. As demonstration, supercoiling dynamics drug-induced DNA break topoisomerases. To rapidly quantify distinct classes behaviors events, developed software platform an automated feature classification pipeline. can be readily adapted for studies broad range complex, multistep enzymatic pathways escaped