Spatio-angular fluorescence microscopy III. Constrained angular diffusion, polarized excitation, and high-NA imaging

We investigate rotational diffusion of fluorescent molecules in angular potential wells, the excitation and subsequent emissions from these diffusing molecules, and the imaging of these emissions with high-NA aplanatic optical microscopes. Although dipole emissions only transmit six low-frequency an...

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Veröffentlicht in:Journal of the Optical Society of America. A, Optics, image science, and vision Optics, image science, and vision, 2020-09, Vol.37 (9), p.1465-1479
Hauptverfasser: Chandler, Talon, Shroff, Hari, Oldenbourg, Rudolf, La Rivière, Patrick
Format: Artikel
Sprache:eng
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Zusammenfassung:We investigate rotational diffusion of fluorescent molecules in angular potential wells, the excitation and subsequent emissions from these diffusing molecules, and the imaging of these emissions with high-NA aplanatic optical microscopes. Although dipole emissions only transmit six low-frequency angular components, we show that angular structured illumination can alias higher-frequency angular components into the passband of the imaging system. We show that the number of measurable angular components is limited by the relationships between three time scales: the rotational diffusion time, the fluorescence decay time, and the acquisition time. We demonstrate our model by simulating a numerical phantom in the limits of fast angular diffusion, slow angular diffusion, and weak potentials.
ISSN:1084-7529
1520-8532
DOI:10.1364/JOSAA.389217