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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

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.…

Point Spread Function (PSF) engineering is used in single emitter localization to measure the emitter position in 3D and possibly other parameters such as the emission color or dipole orientation as well. Advanced PSF models such as spline…

We show that structural information can be extracted from single molecule localization microscopy (SMLM) data. More precisely, we reinterpret SMLM data as the measures of a phaseless optical diffraction tomography system for which the…

Signal Processing · Electrical Eng. & Systems 2023-07-19 Thanh-An Pham , Emmanuel Soubies , Ferréol Soulez , Michael Unser

Single Molecule Localization Microscopy (SMLM) enables the acquisition of high-resolution images by alternating between activation of a sparse subset of fluorescent molecules present in a sample and localization. In this work, the…

Image and Video Processing · Electrical Eng. & Systems 2018-12-17 Arne Bechensteen , Laure Blanc-Féraud , Gilles Aubert

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

Single molecule localization microscopy (SMLM) permits the visualization of cellular structures an order of magnitude smaller than the diffraction limit of visible light, and an accurate, objective evaluation of the resolution of an SMLM…

Molecular fluorescence microscopy is a leading approach to super-resolution and nanoscale imaging in life and material sciences. However, super-resolution fluorescence microscopy is often bottlenecked by system-specific calibrations and…

Single molecule localization microscopy is widely used in biological research for measuring the nanostructures of samples smaller than the diffraction limit. This study uses multifocal plane microscopy and addresses the 3D single molecule…

Computer Vision and Pattern Recognition · Computer Science 2020-01-08 Toshimitsu Aritake , Hideitsu Hino , Shigeyuki Namiki , Daisuke Asanuma , Kenzo Hirose , Noboru Murata

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

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

3D single-pixel imaging (SPI) is a promising imaging technique that can be ffexibly applied to various wavebands. The main challenge in 3D SPI is that the calibration usually requires a large number of standard points as references, which…

Image and Video Processing · Electrical Eng. & Systems 2024-10-11 Xinyue Ma , Chenxing Wang

Among the approaches in three-dimensional (3D) single molecule localization microscopy, there are several point spread function (PSF) engineering approaches, in which depth information of molecules is encoded in 2D images. Usually,the…

Optics · Physics 2024-07-11 Hanzhe Wu , Danni Chen , YiHong Jiand Gan Xiang , Heng Li , Bin Yu , JunLe Qu

We present a method for calibrating the response of a phase-only spatial light modulator in nonlinear microscopy. Our method uses the microscope image itself as calibration measurement and requires no additional hardware components. Our…

Optics · Physics 2025-09-04 Daniël W. S. Cox , Harish Sasikumar , Ivo M. Vellekoop

Super-resolution imaging methods that combine interferometric (z) analysis with single-molecule localization microscopy (iSMLM) have achieved ultra-high 3D precision and contributed to the elucidation of important biological…

Optics · Physics 2025-02-20 Wei Wang , Zengxin Huang , Yilin Wang , Hangfeng Li , Pakorn Kanchanawong

Single-molecule microscopy has become an indispensable tool for biochemical analysis. The capability of characterizing distinct properties of individual molecules without averaging has provided us with a different perspective for the…

Applied Physics · Physics 2020-05-21 Miao-Hsuan Chien , Silvan Schmid

Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging.…

To be able to resolve molecular-clusters it is crucial to access vital informations (such as, molecule density and cluster-size) that are key to understand disease progression and the underlying mechanism. Traditional single-molecule…

Quantitative Methods · Quantitative Biology 2021-01-27 Partha Pratim Mondal

Modulation-enhanced localization microscopy (MELM) has demonstrated significant improvements in both lateral and axial localization precision compared to conventional single-molecule localization microscopy (SMLM). However, lateral…

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