Optical tweezers and multimodality imaging: a platform for dynamic single‐molecule analysis

Abstract

Utilization of single molecule technologies offer researchers the unique opportunity to perform direct real‐time observations and measurements for a plethora of biological mechanisms. Here, we present our efforts to facilitate biologically meaningful discoveries using fully integrated optical tweezers with single‐molecule fluorescence microscopy technology. Correlative fluorescence and optical tweezer microscopy (C‐trap™) is the only technology that truly integrates optical tweezers, confocal/STED microscopy and an advanced microfluidics. The C‐Trap ™ enables live, simultaneous and correlative visualization and manipulation of molecular interactions with sub‐piconewton (pN) force resolution and a kilo‐Hz (to mega‐Hz) temporal resolution, such that even the quickest cellular events can be captured. This technology can be utilized as an easy‐to‐use and stable instrument with the capacity to elucidate biologically meaningful mechanisms. Here we present applications detailing protein (un)folding and conformational changes; DNA‐protein interactions and genome modifications; effects of mechanical stress on DNA/RNA structure; motility of cytoskeletal molecular motors; and protein droplet and aggregation dynamics. These experiments demonstrate technological advances in hybrid single‐molecule methods and the potential for their use in future scientific breakthroughs.