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Single Molecule Localization Microscopy (SMLM) enables the acquisition of high-resolution images by alternating between activation of a sparse subset of fluorescent molecules present in a sample and localization. In this work, the…

Image and Video Processing · Electrical Eng. & Systems 2018-12-17 Arne Bechensteen , Laure Blanc-Féraud , Gilles Aubert

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

MINFLUX (Minimal Photon Flux) is a single-molecule imaging technique capable of resolving fluorophores at a precision of <5 nm. Interpretation of the point patterns generated by this technique presents challenges due to variable emitter…

Applications · Statistics 2026-02-24 Jack Peyton , Benjamin Davis , Emily Gribbin , Daniel Rolfe , Hannah Mitchell

Structured illumination in Single Molecule Localization Microscopy provides new information on the position of molecules and thus improves the localization precision compared to standard localization methods. Here, we used a time-shifted…

We show that structural information can be extracted from single molecule localization microscopy (SMLM) data. More precisely, we reinterpret SMLM data as the measures of a phaseless optical diffraction tomography system for which the…

Signal Processing · Electrical Eng. & Systems 2023-07-19 Thanh-An Pham , Emmanuel Soubies , Ferréol Soulez , Michael Unser

In fluorescence microscopy, Single Molecule Localization Microscopy (SMLM) techniques aim at localizing with high precision high density fluorescent molecules by stochastically activating and imaging small subsets of blinking emitters.…

We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. A 22-fold…

The diffraction of light imposes a fundamental limit on the resolution of light microscopes. This limit can be circumvented by creating and exploiting independent behaviors of the sample at length scales below the diffraction limit. In…

Biological Physics · Physics 2024-06-19 Mohamadreza Fazel , Michael J. Wester

MINFLUX is an emerging super-resolution technology that measures the position of single fluorophores with nanometer precision using fewer photons than any other fluorescence imaging or tracking technique. Here, we derive simple and…

Optics · Physics 2025-01-09 Zach Marin , Jonas Ries

Tracking single fluorescent molecules has offered resolution into dynamic molecular processes at the single-molecule level. This perspective traces the evolution of single-molecule tracking, highlighting key developments across various…

Biological Physics · Physics 2026-02-24 Lance W. Q. Xu , Steve Pressé

Optical super-resolution microscopy is a key technology for structural biology that offers high imaging contrast and live-cell compatibility. Minimal (fluorescence) photons flux microscopy, or MINFLUX, is an emerging super-resolution…

Optics · Physics 2025-08-29 Lukas Scheiderer , Zach Marin , Jonas Ries

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 localization microscopy is a recently developed superresolution imaging technique to visualize structural properties of single cells. The basic principle consists in chemically attaching fluorescent dyes to the molecules,…

In the past years, optical fluorescence microscopy (OFM) made steady progress towards increasing the localisation precision of fluorescent emitters in biological samples. The high precision achieved by these techniques has prompted new…

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) 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 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

Single molecule localization microscopy (SMLM) permits the visualization of cellular structures an order of magnitude smaller than the diffraction limit of visible light, and an accurate, objective evaluation of the resolution of an SMLM…

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

Nanoscale localization of point emitters is critical to several methods in optical fluorescence microscopy, including single-molecule super-resolution imaging and tracking. While the precision of the localization procedure has been the…

Optics · Physics 2020-07-15 Petar N. Petrov , W. E. Moerner
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