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- W3022238431 abstract "Simultaneous measurements of single-molecule positions and orientations provide critical insight into a variety of biological and chemical processes. Various engineered point spread functions (PSFs) have been introduced for measuring the orientation and rotational diffusion of dipole-like emitters, but the widely used Cramér-Rao bound (CRB) only evaluates performance for one specific orientation at a time. Here, we report a performance metric, termed variance upper bound (VUB), that yields a global maximum CRB for all possible molecular orientations, thereby enabling the measurement performance of any PSF to be computed efficiently ( <mml:math xmlns:mml=http://www.w3.org/1998/Math/MathML display=inline> <mml:mrow class=MJX-TeXAtom-ORD> <mml:mo>∼<!-- ∼ --></mml:mo> </mml:mrow> <mml:mn>1000</mml:mn> <mml:mo>×<!-- × --></mml:mo> </mml:math> faster than calculating average CRB). Our VUB reveals that the simple polarized standard PSF provides robust and precise orientation measurements if emitters are near a refractive index interface. Using this PSF, we measure the orientations and positions of Nile red (NR) molecules transiently bound to amyloid aggregates. Our super-resolved images reveal the main binding mode of NR on amyloid fiber surfaces, as well as structural heterogeneities along amyloid fibrillar networks, that cannot be resolved by single-molecule localization alone." @default.
- W3022238431 created "2020-05-13" @default.
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- W3022238431 date "2020-06-04" @default.
- W3022238431 modified "2023-09-30" @default.
- W3022238431 title "Single-molecule orientation localization microscopy for resolving structural heterogeneities between amyloid fibrils" @default.
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- W3022238431 doi "https://doi.org/10.1364/optica.388157" @default.
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