Single-fluorophore orientation determination with multiview polarized illumination : modeling and microscope design

Thumbnail Image
Date
2017-12-01
Authors
Chandler, Talon
Mehta, Shalin B.
Shroff, Hari
Oldenbourg, Rudolf
La Riviere, Patrick J.
Linked Authors
Alternative Title
Date Created
Location
DOI
10.1364/OE.25.031309
Replaced By
Keywords
Abstract
We investigate the use of polarized illumination in multiview microscopes for determining the orientation of single-molecule fluorescence transition dipoles. First, we relate the orientation of single dipoles to measurable intensities in multiview microscopes and develop an information-theoretic metric—the solid-angle uncertainty—to compare the ability of multiview microscopes to estimate the orientation of single dipoles. Next, we compare a broad class of microscopes using this metric—single- and dual-view microscopes with varying illumination polarization, illumination numerical aperture (NA), detection NA, obliquity, asymmetry, and exposure. We find that multi-view microscopes can measure all dipole orientations, while the orientations measurable with single-view microscopes is halved because of symmetries in the detection process. We also find that choosing a small illumination NA and a large detection NA are good design choices, that multiview microscopes can benefit from oblique illumination and detection, and that asymmetric NA microscopes can benefit from exposure asymmetry.
Description
Author Posting. © Optical Society of America, 2017. This article is posted here by permission of Optical Society of America for personal use, not for redistribution. The definitive version was published in Optics Express 25 (2017): 31309-31325, doi:10.1364/OE.25.031309.
Embargo Date
Citation
Optics Express 25 (2017): 31309-31325
Cruises
Cruise ID
Cruise DOI
Vessel Name