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Super-resolution localization microscopy is based on determining the positions of individual fluorescent markers in a sample. The major challenge in reaching an ever higher localization precision lies in the limited number of collected…

Optics · Physics 2020-08-05 Daniel Böning , Franz-Ferdinand Wieser , Vahid Sandoghdar

Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging.…

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

We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and…

Biomolecules · Quantitative Biology 2015-06-24 Zhaokun Zhou , Helen Miller , Adam J. M. Wollman , Mark C. Leake

In single-molecule super-resolution microscopy, engineered point-spread functions (PSFs) are designed to efficiently encode new molecular properties, such as 3D orientation, into complex spatial features captured by a camera. To fully…

Image and Video Processing · Electrical Eng. & Systems 2019-07-16 Hesam Mazidi , Eshan S. King , Oumeng Zhang , Arye Nehorai , Matthew D. Lew

Point-wise localization of individual fluorophores is a critical step in super-resolution microscopy and single particle tracking. Although the methods are limited by the accuracy in localizing individual flourophores, this point-wise…

Biological Physics · Physics 2017-05-08 Martin Lindén , Vladimir Ćurić , Elias Amselem , Johan Elf

The movement of molecules inside living cells is a fundamental feature of biological processes. The ability to both observe and analyse the details of molecular diffusion in vivo at the single molecule and single cell level can add…

Quantitative Methods · Quantitative Biology 2012-11-21 Alex Robson , Kevin Burrage , Mark Leake

We present a single-molecule tool called the CoPro (Concentration of Proteins) method that uses millisecond imaging with convolution analysis, automated image segmentation and super-resolution localization microscopy to generate robust…

Biomolecules · Quantitative Biology 2016-02-17 Adam J. M. Wollman , Mark C. Leake

Super-resolution imaging techniques have largely improved our capabilities to visualize nanometric structures in biological systems. Their application further enables one to potentially quantitate relevant parameters to determine the…

Biological Physics · Physics 2019-12-18 Tina Kosǔta , Marta Cullell-Dalmau , Francesca Cella Zanacchi , Carlo Manzo

FRET-based approaches are a unique tool for sensing the immediate surroundings and interactions of (bio)molecules. FRET imaging and FLIM (Fluorescence Lifetime Imaging Microscopy) enable the visualization of the spatial distribution of…

Single molecule localization microscopy (SMLM) techniques enable imaging biological samples well beyond the diffraction limit of light, but they vary significantly in their spatial and temporal resolutions. High-order statistical analysis…

Biological Physics · Physics 2016-03-15 Shachar Schidorsky , Xiyu Yi , Yair Razvag , Yonatan Golan , Shimon Weiss , Eilon Sherman

Recent technological advances in cutting-edge ultrasensitive fluorescence microscopy have allowed single-molecule imaging experiments in living cells across all three domains of life to become commonplace. Single-molecule live-cell data is…

Biomolecules · Quantitative Biology 2015-04-15 Mark Leake

Single-molecule microscopy has become an indispensable tool for biochemical analysis. The capability of characterizing distinct properties of individual molecules without averaging has provided us with a different perspective for the…

Applied Physics · Physics 2020-05-21 Miao-Hsuan Chien , Silvan Schmid

Fluorescence microscopy is a critical tool across various disciplines, from materials science to biomedical research, yet it is limited by the diffraction limit of resolution. Advanced super-resolution techniques such as localization…

The polarization of light conveys crucial information about the spatial ordering and optical properties of a specimen. However, precise polarization measurement in challenging conditions, including constrained spaces, low light levels, and…

Optics · Physics 2026-03-10 Martin Bielak , Dominik Vašinka , Miroslav Ježek

Fluorescence microscopy is essential in biological and medical research, providing critical insights into cellular structures. However, limited by optical diffraction and background noise, a substantial amount of hidden information is still…

Biological Physics · Physics 2026-04-14 Xiaofeng Zhang , Yongsheng Huang , Jielong Yang , Zhili Wang , Si Chen , Linbo Liu , Xin Ge

The full understanding of cellular functions requires information about protein numbers for various biomolecular assemblies and their dynamics, which can be partly accessed by super-resolution fluorescence microscopy. Yet, many protein…

Biological Physics · Physics 2020-06-17 Kristin S. Grußmayer , Klaus Yserentant , Dirk-Peter Herten

Existing super-resolution microscopy is often constrained by inherent trade-offs between resolution, acquisition speed, phototoxicity, and hardware complexity. Computational post-processing approaches offer a promising alternative, but they…

Super-resolution microscopy is rapidly gaining importance as an analytical tool in the life sciences. A compelling feature is the ability to label biological units of interest with fluorescent markers in living cells and to observe them…

To overcome the physical barriers caused by light diffraction, super-resolution techniques are often applied in fluorescence microscopy. State-of-the-art approaches require specific and often demanding acquisition conditions to achieve…

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