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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

Fluorescent molecules emit light in a dipole radiation pattern that can be used to infer their orientation through defocused fluorescence microscopy. Proper measurement of the orientation requires mathematical modeling of the radiation…

Optics · Physics 2025-08-07 E. Dey , M. Elorza , F. W. Foss , J. J. Gomez Cadenas , B. J. P. Jones

Single molecule localization microscopy is widely used in biological research for measuring the nanostructures of samples smaller than the diffraction limit. This study uses multifocal plane microscopy and addresses the 3D single molecule…

Computer Vision and Pattern Recognition · Computer Science 2020-01-08 Toshimitsu Aritake , Hideitsu Hino , Shigeyuki Namiki , Daisuke Asanuma , Kenzo Hirose , Noboru Murata

A variety of recent imaging techniques are able to beat the diffraction limit in fluorescence microcopy by activating and localizing subsets of the fluorescent molecules in the specimen, and repeating this process until all of the molecules…

Optics · Physics 2010-08-27 Alexander R. Small

Recent efforts to survey the numerous softwares available to perform single molecule tracking (SMT) highlighted a significant dependence of the outcomes on the specific method used, and the limitation encountered by most techniques to…

Quantitative Methods · Quantitative Biology 2019-07-26 Carmine di Rienzo , Paolo Annibale

Localization microscopy is an imaging technique in which the positions of individual nanoscale point emitters (e.g. fluorescent molecules) are determined at high precision from their images. This is the key ingredient in…

Image and Video Processing · Electrical Eng. & Systems 2020-10-01 Elias Nehme , Daniel Freedman , Racheli Gordon , Boris Ferdman , Lucien E. Weiss , Onit Alalouf , Reut Orange , Tomer Michaeli , Yoav Shechtman

We propose a straightforward sample-based technique to calibrate the axial detection in 3D single molecule localization microscopy (SMLM). Using microspheres coated with fluorescent molecules, the calibration curves of PSF-shaping- or…

Single-molecule localization microscopy (SMLM) allows reconstructing biology-relevant structures beyond the diffraction limit by detecting and localizing individual fluorophores -- fluorescent molecules stained onto the observed specimen --…

Computer Vision and Pattern Recognition · Computer Science 2026-03-03 Romain Seailles , Jean-Baptiste Masson , Jean Ponce , Julien Mairal

Single molecule imaging is one of the main target areas of X-ray free electron lasers. It relies on the possibility of orienting the large number of low counting statistics 2D diffraction patterns taken at random orientations of identical…

Atomic and Molecular Clusters · Physics 2012-11-30 Zoltan Jurek , Gyula Faigel

Single particle diffraction imaging experiments at free-electron lasers (FEL) have a great potential for structure determination of reproducible biological specimens that can not be crystallized. One of the challenges in processing the data…

Biological Physics · Physics 2013-10-16 O. M. Yefanov , I. A. Vartanyants

Miniature fluorescence microscopes are a standard tool in systems biology. However, widefield miniature microscopes capture only 2D information, and modifications that enable 3D capabilities increase the size and weight and have poor…

Image and Video Processing · Electrical Eng. & Systems 2020-10-13 Kyrollos Yanny , Nick Antipa , William Liberti , Sam Dehaeck , Kristina Monakhova , Fanglin Linda Liu , Konlin Shen , Ren Ng , Laura Waller

In single molecule localisation super-resolution microscopy the need for repeated image capture limits the imaging speed, while the size of fluorescence probes limits the possible theoretical localisation resolution. Here, we demonstrated a…

Two-dimensional electronic spectroscopy provides information on coupling and energy transfer between excited states on ultrafast timescales. Only recently, incoherent fluorescence detection has made it possible to combine this method with…

Optics · Physics 2025-05-14 Sanchayeeta Jana , Simon Durst , Lucas Ludwig , Markus Lippitz

Tracking single molecules is instrumental for quantifying the transport of molecules and nanoparticles in biological samples, e.g., in brain drug delivery studies. Existing intensity-based localisation methods are not developed for imaging…

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 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

Fluorescent molecules are versatile nanoscale emitters that enable detailed observations of biophysical processes with nanoscale resolution. Because they are well-approximated as electric dipoles, imaging systems can be designed to…

Optics · Physics 2024-03-26 Tingting Wu , Matthew D. Lew

The orientation of luminescent molecules in organic light-emitting diodes (OLEDs) strongly influences device performance. However, our understanding of the factors controlling emitter orientation is limited as current measurements only…

Single particle reconstruction has recently emerged in 3D fluorescence microscopy as a powerful technique to improve the axial resolution and the degree of fluorescent labeling. It is based on the reconstruction of an average volume of a…

Image and Video Processing · Electrical Eng. & Systems 2023-01-24 Thibaut Eloy , Etienne Baudrier , Marine Laporte , Virginie Hamel , Paul Guichard , Denis Fortun

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