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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

We present Full SMS, a multipurpose graphical user interface (GUI)-based software package for analysing single-molecule spectroscopy (SMS) data. SMS typically delivers multiparameter data -- such as fluorescence brightness, lifetime, and…

Biological Physics · Physics 2024-05-01 Joshua L. Botha , Bertus van Heerden , Tjaart P. J. Krüger

Advanced microscopy techniques are essential in biomedical research for visualising and tracking biomolecules within living cells and their compartments. Conventional fluorescence microscopy methods, however, often struggle with accurately…

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

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

Multiplexed immunofluorescence microscopy captures detailed measurements of spatially resolved, multiple biomarkers simultaneously, revealing tissue composition and cellular interactions in situ among single cells. The growing scale and…

Software Engineering · Computer Science 2025-06-03 Fang Liu , Rui He , Andrei Bombin , Ahmad B. Abdallah , Omar Eldaghar , Tommy R. Sheeley , Sam E. Ying , George Zaki

Bright-field microscopy, a cost-effective solution for live-cell culture, is often the only resource available, along with standard CPUs, for many low-budget labs. The inherent challenges of bright-field images -- their noisiness, low…

Quantitative Methods · Quantitative Biology 2025-11-03 Surajit Das , Pavel Zun

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

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…

Single-particle tracking (SPT) grants unprecedented insight into cellular function at the molecular scale [1]. Throughout the cell, the movement of single-molecules is generally heterogeneous and complex. Hence, there is an imperative to…

Quantitative Methods · Quantitative Biology 2015-06-24 Mohamed El Beheiry , Maxime Dahan , Jean-Baptiste Masson

We review methods of data analysis for biophysical data with a special emphasis on single molecule applications. Our review is intended for anyone, from student to established researcher. For someone just getting started, we focus on…

Biological Physics · Physics 2016-11-24 Meysam Tavakoli , J. Nicholas Taylor , Chun-Biu Li , Tamiki Komatsuzaki , Steve Pressé

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

An improved analysis for single particle imaging (SPI) experiments, using the limited data, is presented here. Results are based on a study of bacteriophage PR772 performed at the AMO instrument at the Linac Coherent Light Source (LCLS) as…

Processing of medical images such as MRI or CT presents unique challenges compared to RGB images typically used in computer vision. These include a lack of labels for large datasets, high computational costs, and metadata to describe the…

Image and Video Processing · Electrical Eng. & Systems 2021-08-06 Fernando Pérez-García , Rachel Sparks , Sébastien Ourselin

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

Optical detection and spectroscopy of single molecules has become an indispensable tool in biological imaging and sensing. Its success is based on fluorescence of organic dye molecules under carefully engineered laser illumination. In this…

Chemical Physics · Physics 2011-01-18 Ilja Gerhardt , Lijian Mai , Antia Lamas-Linares , Christian Kurtsiefer

The Linac Coherent Light Source (LCLS) is an X- ray free electron laser (XFEL) facility enabling the study of the structure and dynamics of single macromolecules. A major upgrade will bring the repetition rate of the X-ray source from 120…

Computational Physics · Physics 2021-09-14 Hsing-Yin Chang , Elliott Slaughter , Seema Mirchandaney , Jeffrey Donatelli , Chun Hong Yoon

Determining molecular abundances in astrophysical environments is crucial for interpreting observational data and constraining physical conditions in these regions. Chemical modelling tools are essential for simulating the complex processes…

Instrumentation and Methods for Astrophysics · Physics 2025-09-17 Luke Keyte , Jason Ran

We present a simple and robust technique to extract kinetic rate models and thermodynamic quantities from single molecule time traces. SMACKS (Single Molecule Analysis of Complex Kinetic Sequences) is a maximum likelihood approach that…

Quantitative Methods · Quantitative Biology 2022-03-10 Sonja Schmid , Markus Götz , Thorsten Hugel

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
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