To characterize the molecular structure and function of the actomyosin motor and its modulation by myosin binding protein-C (MyBP-C), we have implemented an array of single molecule biophysical techniques that can measure the force and motion of a single molecular motor or any protein-protein interaction. Examples of these techniques are the laser trap and super-resolution STORM (STochastic Optical Reconstruction Microscopy) microscopy. Novel biophysical techniques are combined to address interesting research problems ranging from heart disease to the ability of myosin motors to navigate the complex cytoskeleton within cells during cargo transport.