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Abstract
Single-molecule localization and orientation microscopy (SMOLM) measures both the orientations and locations of single molecules; this technology has pushed precision towards the physical limit. My thesis is aimed at improving the precision of orientation measurements by finding the optimal excitation modulation direction. For the multi-view reflector (MVR) microscope, we developed both linear and circular polarization schemes. Among linear schemes, L = 4 achieves the most uniform coverage per frame. When having the same number of excitation frames, the optimized L = 6 scheme outperforms the theoretically isotropic design, termed equal pumping, by having higher orientation measurement precision. Extending the optimization to four engineered dipole-spread function (DSF) microscopes–double helix, pixOL, vortex, and CHIDO–we find optimal frame counts of L = 6, 5, 5, and 6, respectively, with the largest Cramér-Rao bound (CRB) reductions occurring at polar angles θ where each DSF is inherently less sensitive. These results provide practical design guidelines for SMOLM modulated by excitation polarization across a wide range of DSFs and experimental conditions.
Document Type
Article
Class Name
Electrical and Systems Engineering Undergraduate Research
Language
English (en)
Date of Submission
5-7-2026
Recommended Citation
Nie, Kaizhi A., "Excitation Modulation Optimization on Single-Molecule Orientation Microscopy" (2026). Electrical and Systems Engineering Undergraduate and Graduate Research. 61.
https://openscholarship.wustl.edu/eseundergraduate_research/61