Engineering Commons^™

Measuring Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang

McKelvey School of Engineering Theses & Dissertations

Single molecules have become a powerful tool for biophysicists since they were first optically detected 28 years ago. Understanding molecular orientation can not only improve the accuracy of single-molecule localization, but it can also provide insight into biochemical behaviors at the nanoscale. In this thesis, I present a method to measure the molecular orientation and rotational mobility of single-molecule emitters by designing and implementing a tri-spot point spread function. The point spread function is designed so that it is capable of measuring all degrees of freedom related to molecular orientation and rotational mobility. Its design is optimized by maximizing the …