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

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    oe-25-25-31309.pdf (20.05Mb)
    Date
    2017-12-01
    Author
    Chandler, Talon  Concept link
    Mehta, Shalin B.  Concept link
    Shroff, Hari  Concept link
    Oldenbourg, Rudolf  Concept link
    La Riviere, Patrick J.  Concept link
    Metadata
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    Citable URI
    https://hdl.handle.net/1912/9446
    As published
    https://doi.org/10.1364/OE.25.031309
    DOI
    10.1364/OE.25.031309
    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.
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    Suggested Citation
    Optics Express 25 (2017): 31309-31325
     
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