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We demonstrate sub-wavelength sectioning on biological samples with a conventional confocal microscope. This optical sectioning is achieved by the phenomenon of supercritical angle fuorescence, wherein only a fluorophore next to the…

We introduce the basic elements of a spatio-angular theory of fluorescence microscopy, providing a unified framework for analyzing systems that image single fluorescent dipoles and ensembles of overlapping dipoles that label biological…

Precisely measuring the three-dimensional position and orientation of individual fluorophores is challenging due to the substantial photon shot noise in single-molecule experiments. Facing this limited photon budget, numerous techniques…

Optics · Physics 2021-02-02 Oumeng Zhang , Matthew D. Lew

The back-focal plane (BFP) of a high-numerical aperture objective contains the fluoro-phore radiation pattern, which encodes information about the axial fluorophore position, molecular orientation and the local refractive index of the…

Optical fluorescence imaging is capable of measuring both the spatial and rotational dynamics of single molecules. However, unavoidable measurement noise will result in inaccurate estimates of rotational dynamics, causing a molecule to…

Optics · Physics 2019-05-22 Oumeng Zhang , Matthew D. Lew

Our work aims at using quantitative imaging tools to complement the limitation of noise encountered by high resolution fluorescence microscopy methods. Several cycles of fluorophore activation, imaging and deactivation produce a sequence of…

Quantitative Methods · Quantitative Biology 2012-02-03 Juliane Liepe , Federico Felizzi , Agata Pernus , Maria Hanulova

Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background…

Optics · Physics 2021-02-02 Oumeng Zhang , Matthew D. Lew

Point spread function (PSF) plays an essential role in image reconstruction. In the context of confocal microscopy, optical performance degrades towards the edge of the field of view as astigmatism, coma and vignetting. Thus, one should…

Computer Vision and Pattern Recognition · Computer Science 2017-08-01 Anna Jezierska , Hugues Talbot , Jean-Christophe Pesquet , Gilbert Engler

Single-shot volumetric fluorescence (SVF) imaging offers a significant advantage over traditional imaging methods that require scanning across multiple axial planes as it can capture biological processes with high temporal resolution. The…

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.…

We present a new technique for monitoring microlensing activity even in highly crowded fields, and use this technique to place limits on low-mass MACHOs in the haloes of M31 and the Galaxy. Unlike present Galactic microlensing surveys, we…

Astrophysics · Physics 2009-09-28 Austin B. Tomaney , Arlin P. S. Crotts

The main advantage of two-photon fluorescence confocal microscopy is the low absorption obtained with live tissues at the wavelengths of operation. However, the resolution of two-photon fluorescence confocal microscopes is lower than in the…

Biological Physics · Physics 2009-11-13 N. Sandeau , H. Giovannini

Super-resolution mapping of the 3D orientation of fluorophores reveals the alignment of biological structures where the fluorophores are tightly attached, and thus plays a vital role in studying the organization and dynamics of…

Optics · Physics 2024-01-23 Xiaowei Liu , Mingwei Tang , Ning Zhou , Chenlei Pang , Zhong Wen , Xu Liu , Qing Yang

Fluorescence microscopy is an important and extensively utilised tool for imaging biological systems. However, the image resolution that can be obtained has a limit as defined through the laws of diffraction. Demand for improved resolution…

Biological Physics · Physics 2007-08-27 James H. Rice

A new far-field optical microscopy technique capable of reaching nanometer-scale resolution has been developed recently using the in-plane image magnification by surface plasmon polaritons. This microscopy is based on the optical properties…

Strongly Correlated Electrons · Physics 2009-11-10 Igor I. Smolyaninov

We suggest a scheme to manipulate paraxial diffraction by utilizing the dependency of a four-wave mixing process on the relative angle between the light fields. A microscopic model for four-wave mixing in a Lambda-type level structure is…

Optics · Physics 2015-06-18 Itay Katzir , Amiram Ron , Ofer Firstenberg

The fields of view of Extremely Large Telescopes will contain vast numbers of spatial sampling elements (spaxels) as their Adaptive Optics systems approach the diffraction limit over wide fields. Since this will exceed the detection…

Instrumentation and Methods for Astrophysics · Physics 2015-05-13 G. J. Murray , J. R. Allington-Smith

Fluorescence fluctuation spectroscopy is applied to study molecules, passing through a small observation volume, usually subjected to diffusive or convective motion in liquid phase. We suggest that such a technique could be used to measure…

Fluorescent observation of cells generally suffers from the limited axial resolution due to the elongated point spread function of the microscope optics. Consequently, three-dimensional imaging results in axial resolution being several…

Fluorescence detection, either involving propagating or near-field emission, is widely being used in spectroscopy, sensing and microscopy. Total internal reflection fluorescence (TIRF) confines fluorescence excitation by an evanescent…

Quantitative Methods · Quantitative Biology 2020-06-24 Martin Oheim , Adi Salomon , Maia Brunstein
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