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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…

生物大分子 · 定量生物学 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…

生物物理 · 物理学 2023-01-25 Jack W Shepherd , Alex L Payne-Dwyer , Ji-Eun Lee , Aisha Syeda , Mark C Leake

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…

生物物理 · 物理学 2015-05-18 Shawn H. DeCenzo , Michael C. DeSantis , Y. M. Wang

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…

生物物理 · 物理学 2024-05-01 Joshua L. Botha , Bertus van Heerden , Tjaart P. J. Krüger

Super-resolution microscopy is rapidly gaining importance as an analytical tool in the life sciences. A compelling feature is the ability to label biological units of interest with fluorescent markers in living cells and to observe them…

Fluorescence lifetime imaging microscopy (FLIM) provides detailed information about molecular interactions and biological processes. A major bottleneck for FLIM is image resolution at high acquisition speeds, due to the engineering and…

图像与视频处理 · 电气工程与系统科学 2024-04-23 Valentin Kapitány , Areeba Fatima , Vytautas Zickus , Jamie Whitelaw , Ewan McGhee , Robert Insall , Laura Machesky , Daniele Faccio

Single-molecule spectroscopy (SMS) is an exceptionally sensitive technique, but its inherently limited photon budget produces noisy data that can readily lead to subjective analyses, fitting errors, and reduced statistical power, obscuring…

生物物理 · 物理学 2026-03-19 Michael Lovemore , Joshua Botha , Bertus van Heerden , Tjaart Kruger

Super-resolution Structured Illumination Microscopy (SR-SIM) enables fluorescence microscopy beyond the diffraction limit at high frame rates. Compared to other super-resolution microscopy techniques, the low photon fluence used in SR-SIM…

We report, for the first time, a multi-confocal Fluorescence Correlation Spectroscopy (mFCS) technique which allows parallel measurements at different locations, by combining a Spatial Light Modulator (SLM), with an Electron Multiplying-CCD…

As camera pixel arrays have grown larger and faster, and optical microscopy techniques ever more refined, there has been an explosion in the quantity of data acquired during routine light microcopy. At the single-molecule level, analysis…

生物大分子 · 定量生物学 2021-07-07 Jack W Shepherd , Ed J Higgins , Adam J M Wollman , Mark C Leake

Endoscopic optical imaging using a single multimode fiber (MMF) has emerged as a promising approach for highly compact, minimally invasive, and high-resolution imaging. Unlike conventional fiber bundles, MMF-based endomicroscopes exploit…

Scanning fluorescence correlation spectroscopy (SFCS) with a scan path perpendicular to the membrane plane was introduced to measure diffusion and interactions of fluorescent components in free standing biomembranes. Using a confocal laser…

生物大分子 · 定量生物学 2018-06-04 Paul Müller , Petra Schwille , Thomas Weidemann

Compressive Macroscopic Fluorescence Lifetime Imaging (MFLI) is a novel technical implementation that enables monitoring multiple molecular interactions in macroscopic scale. Especially, we reported recently on the development of a…

医学物理 · 物理学 2018-02-23 Ruoyang Yao , Marien Ochoa , Xavier Intes , Pingkun Yan

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…

细胞行为 · 定量生物学 2009-11-10 R. V. Krishnan , Eva Biener , Jian-Hua Zhang , Robert Heckel , Brian Herman

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…

Due to its specificity, fluorescence microscopy (FM) has become a quintessential imaging tool in cell biology. However, photobleaching, phototoxicity, and related artifacts continue to limit FM's utility. Recently, it has been shown that…

Fluorescence spectroscopy is an image correlation technique to analyze and characterize the molecular dynamics from a sequence of fluorescence images. Many image correlation techniques have been developed for different applications [1]. But…

定量方法 · 定量生物学 2017-10-24 C. Chen , P. Paul-Gilloteaux , T. Vignaud , J. Salamero , F. Waharte

Fluorescence microscopy is a widely used method among cell biologists for studying the localization and co-localization of fluorescent protein. For microbial cell biologists, these studies often include tedious and time-consuming manual…

计算机视觉与模式识别 · 计算机科学 2020-09-29 Jonathan Reiner , Guy Azran , Gal Hyams

Novel techniques are currently being developed and established for the accurate chemical analysis and detection of single cancer cells, single embryos and single seeds by Fourier Transform Near Infrared (FT-NIR) Microspectroscopy, Fourier…

组织与器官 · 定量生物学 2007-05-23 I. C. Baianu , D. Costescu , N. E. Hofmann , S. S. Korban

Fluorescence imaging is the most widely used method for unveiling the molecular composition of biological specimens. However, the weak optical emission of fluorescent probes and the tradeoff between imaging speed and sensitivity is…

光学 · 物理学 2015-06-15 Eric D. Diebold , Brandon W. Buckley , Daniel R. Gossett , Bahram Jalali
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