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Full-Text Articles in Physics
Imaging The Three-Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri-Spot Point Spread Function, Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew
Imaging The Three-Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri-Spot Point Spread Function, Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
Fluorescence photons emitted by single molecules contain rich information regarding their rotational motions, but adapting single-molecule localization microscopy (SMLM) to measure their orientations and rotational mobilities with high precision remains a challenge. Inspired by dipole radiation patterns, we design and implement a Tri-spot point spread function (PSF) that simultaneously measures the three-dimensional orientation and the rotational mobility of dipole-like emitters across a large field of view. We show that the orientation measurements done using the Tri-spot PSF are sufficiently accurate to correct the anisotropy-based localization bias, from 30 nm to 7 nm, in SMLM. We further characterize the emission anisotropy …
Measuring 3d Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang, Tianben Ding, Jin Lu, Hesam Mazidi, Matthew D. Lew
Measuring 3d Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang, Tianben Ding, Jin Lu, Hesam Mazidi, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
We 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. It can measure all degrees of freedom related to molecular orientation and rotational mobility. Its design is optimized by maximizing the theoretical limit of its measurement precision. We evaluate the precision and accuracy of the Tri-spot PSF by measuring the orientation and effective rotational mobility of single fluorescent molecules embedded in a polymer matrix.