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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…

Resonant Energy Transfer (RET) from an optically excited donor molecule (D) to a non-excited acceptor molecule (A) residing nearby is widely used to detect molecular interactions in living cells. Stoichiometric information, such as the…

Single-molecule narrowfield microscopy is a versatile tool to investigate a diverse range of protein dynamics in live cells and has been extensively used in bacteria. Here, we describe how these methods can be extended to larger eukaryotic,…

Subcellular Processes · Quantitative Biology 2016-06-09 Adam J. M. Wollman , Mark C. Leake

Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system's spatial resolution. Here we report on…

Quantitative Methods · Quantitative Biology 2015-05-13 Michael C. DeSantis , Shawn H. DeCenzo , Je-Luen Li , Y. M. Wang

In example-based super-resolution, the function relating low-resolution images to their high-resolution counterparts is learned from a given dataset. This data-driven approach to solving the inverse problem of increasing image resolution…

Image and Video Processing · Electrical Eng. & Systems 2018-12-05 Alexander Robey , Vidya Ganapati

We present an ultra-fast, precise, parameter-free method, which we term Deep-STORM, for obtaining super-resolution images from stochastically-blinking emitters, such as fluorescent molecules used for localization microscopy. Deep-STORM uses…

Optics · Physics 2018-05-03 Elias Nehme , Lucien E. Weiss , Tomer Michaeli , Yoav Shechtman

Optoplasmonic methods capable of single protein detection so far rely on analyte immobilization in order to facilitate detection [1-6]. These detection schemes, even if they facilitate transient single-molecule detection [7,8] via…

Optics · Physics 2022-01-21 M. D. Baaske , N. Asgari , D. Punj , M. Orrit

The importance of spin-correlated radical pairs in biology is increasingly recognized, with roles in biological effects of weak magnetic fields and emerging quantum spin-based biomedical applications. Fluorescence microscopy offers…

Biological Physics · Physics 2025-10-08 Noboru Ikeya , Jonathan R. Woodward

Surface molecules, distributed in diverse patterns and clusters on cell membranes, influence vital functions of living cells. It is therefore important to understand their molecular surface organisation under different physiological and…

The random motion of molecules in living cells has consistently been reported to deviate from standard Brownian motion, a behavior coined as ``anomalous diffusion''. Fluorescence Correlation Spectroscopy (FCS) is a powerful method to…

Quantitative Methods · Quantitative Biology 2024-07-18 Nathan Quiblier , Jan-Michael Rye , Pierre Leclerc , Henri Truong , Abdelkrim Hannou , Laurent Héliot , Hugues Berry

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

Fluorescence lifetime imaging microscopy (FLIM) is a powerful quantitative technique that provides metabolic and molecular contrast, offering strong translational potential for label-free, real-time diagnostics. However, its clinical…

Computer Vision and Pattern Recognition · Computer Science 2025-12-19 Paloma Casteleiro Costa , Parnian Ghapandar Kashani , Xuhui Liu , Alexander Chen , Ary Portes , Julien Bec , Laura Marcu , Aydogan Ozcan

Ultrasound localization microscopy has enabled super-resolution vascular imaging through precise localization of individual ultrasound contrast agents (microbubbles) across numerous imaging frames. However, analysis of high-density regions…

Signal Processing · Electrical Eng. & Systems 2018-12-14 Ruud J. G. van Sloun , Oren Solomon , Matthew Bruce , Zin Z. Khaing , Hessel Wijkstra , Yonina C. Eldar , Massimo Mischi

Far-field super-resolution fluorescence microscopy has been rapidly developed for applications ranging from cell biology to nanomaterials. However, it remains a significant challenge to achieve super-resolution imaging at depth in opaque…

Optics · Physics 2024-05-01 Tengfei Wu , YoonSeok Baek , Fei Xia , Sylvain Gigan , Hilton B. de Aguiar

Polarized light microscopy, as a contrast-enhancing technique for optically anisotropic materials, is a method well suited for the investigation of a wide variety of effects in solid-state physics, as for example birefringence in crystals…

Instrumentation and Detectors · Physics 2017-12-19 Matthias Lange , Stefan Guénon , Felix Lever , Reinhold Kleiner , Dieter Koelle

We present high resolution two-photon excitation microscopy studies combining two-photon fluorescence (TPF) and second harmonic generation (SHG) in order to probe orientational distributions of molecular ensembles at room temperature. A…

Soft Condensed Matter · Physics 2007-05-23 Veronique Le Floc'h , Sophie Brasselet , Jean-Francois Roch , Joseph Zyss

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

Understanding and extracting the patterns of microscopy images has been a major challenge in the biomedical field. Although trained scientists can locate the proteins of interest within a human cell, this procedure is not efficient and…

Computer Vision and Pattern Recognition · Computer Science 2020-06-09 Hanke Chen

Quality control in molecular optical sectioning microscopy is indispensable for transforming acquired digital images from qualitative descriptions to quantitative data. Although numerous tools, metrics, and phantoms have been developed,…

Single-molecule approaches to biology offer a powerful new vision to elucidate the mechanisms that underpin the functioning of living cells. However, conventional optical single molecule spectroscopy techniques such as F\"orster…

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