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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

Localization of single fluorescent molecules is key for physicochemical and biophysical measurements such as single-molecule tracking and super-resolution imaging by single-molecule localization microscopy (SMLM). Recently a series of…

Optics · Physics 2021-09-06 Luciano A. Masullo , Lucía F. Lopez , Fernando D. Stefani

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.…

Super-resolution microscopy has catalyzed valuable insights into the sub-cellular, mechanistic details of many different biological processes across a wide range of cell types. Fluorescence polarization spectroscopy tools have also enabled…

Biological Physics · Physics 2023-01-25 Jack W Shepherd , Alex L Payne-Dwyer , Ji-Eun Lee , Aisha Syeda , Mark C Leake

Super-resolution microscopy has revolutionized optical fluorescence imaging by improving 3D resolution by 1-2 orders of magnitude. While different methods can successfully increase the resolution, all methods share significant differences…

Biological Physics · Physics 2015-01-26 Thomas Pengo , Nicolas Olivier , Suliana Manley

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

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

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

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…

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

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

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

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

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

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

A long-standing challenge in multiple-particle-tracking is the accurate and precise 3D localization of individual particles at close proximity. One established approach for snapshot 3D imaging is point-spread-function (PSF) engineering, in…

Image and Video Processing · Electrical Eng. & Systems 2020-10-01 Elias Nehme , Boris Ferdman , Lucien E. Weiss , Tal Naor , Daniel Freedman , Tomer Michaeli , Yoav Shechtman

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

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.…

Colocalization is a powerful tool to study the interactions between fluorescently labeled molecules in biological fluorescence microscopy. However, existing techniques for colocalization analysis have not undergone continued development…

Methodology · Statistics 2018-02-14 Shulei Wang , Ellen T. Arena , Kevin W. Eliceiri , Ming Yuan
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