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Over the last decade, single-molecule optical microscopy has become the gold-standard approach to decipher complex molecular processes in cellular environments. [1-3] Single-molecule fluorescence microscopy has several advantages such as…

Biological Physics · Physics 2017-11-15 Edakkattuparambil Sidharth Shibu , Nadezda Varkentina , Laurent Cognet , Brahim Lounis

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.…

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

Measuring subdiffraction separations between single fluorescent particles is important for biological, nano-, and medical-technology studies. Major challenges include (i) measuring changing molecular separations with high temporal…

Biological Physics · Physics 2015-05-18 Shawn H. DeCenzo , Michael C. DeSantis , Y. M. Wang

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

Fluorescence Correlation Spectroscopy (FCS) is a powerful single-molecule technique which allows for measuring motion (diffusion, flow), concentration, and molecular interaction kinetics of fluorescent molecules from picomolar to micromolar…

Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the universal…

This study addresses the critical need for high signal-to-noise ratio in optical detection methods for biological sample discrimination under low-photon-flux conditions to ensure accuracy without compromising sample integrity. We explore…

Optics · Physics 2024-01-22 L. Zhang , V. R. Besaga , P. Rühl , C. Zou , S. H. Heinemann , Y. Wang , F. Setzpfandt

Confocal microscopy of fluorescent labeled particles has been used to study the dynamical and structural properties of colloidal and granular matter in real space. Localization algorithms allow for a fully automatized determination of the…

Soft Condensed Matter · Physics 2011-12-08 Marcel Roth , Michael Franzmann , Maria D'Acunzi , Max Kreiter , Günter K. Auernhammer

Single-shot fluorescence imaging techniques have gained increasing interest in recent years due to their ability to rapidly capture complex biological data without the need for extensive scanning. In this letter, we introduce polarized…

Optics · Physics 2025-01-31 Oumeng Zhang , Changhuei Yang

Single-molecule detection enables direct observation of individual biomolecular events, providing mechanistic insights into biological processes and offering a powerful tool for disease diagnostics. However, the fundamental scale mismatch…

Optics · Physics 2025-10-14 Guifeng Li , Chaoyang Gong

We combine stroboscopic laser excitation with stochastic photoactivation and super-resolution fluorescence imaging. This makes it possible to record hundreds of diffusion trajectories of small protein molecules in single bacterial cells…

Quantitative Methods · Quantitative Biology 2010-03-11 Brian P. English , Arash Sanamrad , Stoyan Tankov , Vasili Hauryliuk , Johan Elf

The advent of single molecule microscopy has revolutionized biological investigations by providing a powerful tool for the study of intercellular and intracellular trafficking processes of protein molecules which was not available before…

Signal Processing · Electrical Eng. & Systems 2020-11-26 Milad R. Vahid , Bernard Hanzon , Raimund J. Ober

As proof-of-principle for generating superresolution structural information from DNA we applied a method of localization microscopy utilizing photoblinking comparing intercalating dye YOYO-1 against minor groove binding dye SYTO-13, using a…

Biomolecules · Quantitative Biology 2015-04-15 Helen Miller , Zhaokun Zhou , Adam J. M. Wollman , Mark C. Leake

We report the cell biological applications of a recently developed multiphoton fluorescence lifetime imaging microscopy system using a streak camera (StreakFLIM). The system was calibrated with standard fluorophore specimens and was shown…

Cell Behavior · Quantitative Biology 2009-11-10 R. V. Krishnan , Eva Biener , Jian-Hua Zhang , Robert Heckel , Brian Herman

An interferometric fluorescent microscope and a novel theoretic image reconstruction approach were developed and used to obtain super-resolution images of live biological samples and to enable dynamic real time tracking. The tracking…

The combination of different imaging modalities into single imaging platforms has a strong potential in biomedical sciences since it permits the analysis of complementary properties of the target sample. Here, we report on an extremely…

Optics · Physics 2025-01-17 David Alonso , Javier Garcia , Vicente Mico

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…

Single-shot volumetric fluorescence (SVF) imaging offers a significant advantage over traditional imaging methods that require scanning across multiple axial planes as it can capture biological processes with high temporal resolution. The…

Precise measurement of multi-parameters of ultrafast lasers is vital both in scientific investigations and technical applications, such as, optical field manipulation, pulse shaping, sample characteristics test, and biomedical imaging.…