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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

Fluorescence microscopy has enabled a dramatic development in modern biology by visualizing biological organisms with micrometer scale resolution. However, due to the diffraction limit, sub-micron/nanometer features are difficult to…

Image and Video Processing · Electrical Eng. & Systems 2021-03-10 Varun Mannam , Yide Zhang , Xiaotong Yuan , Scott Howard

Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background…

Optics · Physics 2021-02-02 Oumeng Zhang , Matthew D. Lew

To overcome the physical barriers caused by light diffraction, super-resolution techniques are often applied in fluorescence microscopy. State-of-the-art approaches require specific and often demanding acquisition conditions to achieve…

In some super-resolution techniques, adjacent points are illuminated at different times. Thereby, their locations and light intensities can be detected even if the images are very blurred due to diffraction. According to conventional…

Image and Video Processing · Electrical Eng. & Systems 2019-12-10 Edward Y. Sheffield

Our work aims at using quantitative imaging tools to complement the limitation of noise encountered by high resolution fluorescence microscopy methods. Several cycles of fluorophore activation, imaging and deactivation produce a sequence of…

Quantitative Methods · Quantitative Biology 2012-02-03 Juliane Liepe , Federico Felizzi , Agata Pernus , Maria Hanulova

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…

Fluorescence microscopy is a critical tool across various disciplines, from materials science to biomedical research, yet it is limited by the diffraction limit of resolution. Advanced super-resolution techniques such as localization…

Fluorescence microscopy is an important and extensively utilised tool for imaging biological systems. However, the image resolution that can be obtained has a limit as defined through the laws of diffraction. Demand for improved resolution…

Biological Physics · Physics 2007-08-27 James H. Rice

Diffraction-limited imaging in epi-fluorescence microscopy remains a challenge when sample aberrations are present or when the region of interest rests deep within an inhomogeneous medium. Adaptive optics is an attractive solution albeit…

Two-dimensional electronic spectroscopy provides information on coupling and energy transfer between excited states on ultrafast timescales. Only recently, incoherent fluorescence detection has made it possible to combine this method with…

Optics · Physics 2025-05-14 Sanchayeeta Jana , Simon Durst , Lucas Ludwig , Markus Lippitz

High-throughput biological imaging is often constrained by a trade-off between acquisition speed and image quality. Fast imaging modalities, such as wide-field fluorescence microscopy, enable large-scale data acquisition but suffer from…

Image and Video Processing · Electrical Eng. & Systems 2026-04-20 Dominik Panek , Carina Rząca , Maksymilian Szczypior , Joanna Sorysz , Krzysztof Misztal , Zbigniew Baster , Zenon Rajfur

We introduce a procedure to automatically count and locate the fluorescent particles in a microscopy image. Our procedure employs an approximate likelihood estimator derived from a Poisson random field model for photon emission. Estimates…

Applications · Statistics 2010-11-10 John Hughes , John Fricks , William Hancock

In single molecule localisation super-resolution microscopy the need for repeated image capture limits the imaging speed, while the size of fluorescence probes limits the possible theoretical localisation resolution. Here, we demonstrated a…

Improving the resolution of fluorescence microscopy beyond the diffraction limit can be achievedby acquiring and processing multiple images of the sample under different illumination conditions.One of the simplest techniques, Random…

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…

Single-molecule localization fluorescence microscopy constructs super-resolution images by sequential imaging and computational localization of sparsely activated fluorophores. Accurate and efficient fluorophore localization algorithms are…

Image and Video Processing · Electrical Eng. & Systems 2020-07-21 Artur Speiser , Lucas-Raphael Müller , Ulf Matti , Christopher J. Obara , Wesley R. Legant , Jonas Ries , Jakob H. Macke , Srinivas C. Turaga

Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique in biomedical research that uses the fluorophore decay rate to provide additional contrast in fluorescence microscopy. However, at present, the calculation, analysis,…

Image and Video Processing · Electrical Eng. & Systems 2021-06-09 Varun Mannam , Yide Zhang , Xiaotong Yuan , Cara Ravasio , Scott S. Howard

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

The article demonstrates some less known principles of image build-up in diffractive microscopy and their usage in analysis unravelling the smallest localized information about the original object - an electromagnetic centroid. In…

Medical Physics · Physics 2016-12-15 Renata Rychtarikova , Georg Steiner , Michael B. Fischer , Dalibor Stys
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