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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

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

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

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

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

The point spread function (PSF) is fundamental to any type of microscopy, most importantly so for single-molecule localization techniques, where the exact PSF shape is crucial for precise molecule localization at the nanoscale. However,…

Super-resolution localization microscopy is based on determining the positions of individual fluorescent markers in a sample. The major challenge in reaching an ever higher localization precision lies in the limited number of collected…

Optics · Physics 2020-08-05 Daniel Böning , Franz-Ferdinand Wieser , Vahid Sandoghdar

Nanoparticles (NPs) have proven their applicability in biosensing, drug delivery, and photo-thermal therapy, but their performance depends critically on the distribution and number of functional groups on their surface. When studying…

Fluorescence microscopy is essential in biological and medical research, providing critical insights into cellular structures. However, limited by optical diffraction and background noise, a substantial amount of hidden information is still…

Biological Physics · Physics 2026-04-14 Xiaofeng Zhang , Yongsheng Huang , Jielong Yang , Zhili Wang , Si Chen , Linbo Liu , Xin Ge

Localization microscopy is an imaging technique in which the positions of individual nanoscale point emitters (e.g. fluorescent molecules) are determined at high precision from their images. This is the key ingredient in…

Image and Video Processing · Electrical Eng. & Systems 2020-10-01 Elias Nehme , Daniel Freedman , Racheli Gordon , Boris Ferdman , Lucien E. Weiss , Onit Alalouf , Reut Orange , Tomer Michaeli , Yoav Shechtman

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…

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

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…

Optimal error estimation is key to achieve accurate photometry and astrometry. Stellar fluxes and positions in high angular resolution images are typically measured with PSF fitting routines, such as StarFinder. However, the formal…

Instrumentation and Methods for Astrophysics · Physics 2022-08-12 E. Gallego-Cano , R. Schödel , A. T. Gallego-Calvente , A. M. Ghez

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

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

The common assumption that precision is the limit of accuracy in localization microscopy and the typical absence of comprehensive calibration of optical microscopes lead to a widespread issue - overconfidence in measurement results with…

Instrumentation and Detectors · Physics 2021-06-21 Craig R. Copeland , Jon Geist , Craig D. McGray , Vladimir A. Aksyuk , J. Alexander Liddle , B. Robert Ilic , Samuel M. Stavis

Single molecule localization microscopy is a recently developed superresolution imaging technique to visualize structural properties of single cells. The basic principle consists in chemically attaching fluorescent dyes to the molecules,…

Traceability to the International System of Units (SI) is fundamental to measurement accuracy and reliability. In this study, we demonstrate subnanometer traceability of localization microscopy, establishing a metrological foundation for…

To ensure high quality outputs, it is important to quantify the epistemic uncertainty of diffusion models. Existing methods are often unreliable because they mix epistemic and aleatoric uncertainty. We introduce a method based on Fisher…

Machine Learning · Statistics 2026-02-18 Aditi Gupta , Raphael A. Meyer , Yotam Yaniv , Elynn Chen , N. Benjamin Erichson
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