scholarly journals Observation of live chromatin dynamics in cells via 3D localization microscopy using Tetrapod point spread functions

2017 ◽  
Vol 8 (12) ◽  
pp. 5735 ◽  
Author(s):  
Yoav Shechtman ◽  
Anna-Karin Gustavsson ◽  
Petar N. Petrov ◽  
Elisa Dultz ◽  
Maurice Y. Lee ◽  
...  
2019 ◽  
Vol 25 (S2) ◽  
pp. 1232-1233
Author(s):  
Jason T. Martineau ◽  
Rajesh Menon ◽  
Erik M. Jorgensen ◽  
Jordan Gerton

2020 ◽  
Author(s):  
Koen J.A. Martens ◽  
Abbas Jabermoradi ◽  
Suyeon Yang ◽  
Johannes Hohlbein

The point spread function (PSF) of single molecule emitters can be engineered in the Fourier plane to encode three-dimensional localization information, creating double-helix, saddle-point or tetra-pod PSFs. Here, we describe and assess adaptations of the phasor-based single-molecule localization microscopy (pSMLM) algorithm to localize single molecules using these PSFs with sub-pixel accuracy. For double-helix, pSMLM identifies the two individual lobes and uses their relative rotation for obtaining z-resolved localizations, while for saddle-point or tetra-pod, a novel phasor-based deconvolution approach is used. The pSMLM software package delivers similar precision and recall rates to the best-in-class software package (SMAP) at signal-to-noise ratios typical for organic fluorophores. pSMLM substantially improves the localization rate by a factor of 2 - 4x on a standard CPU, with 1-1.5·104 (double-helix) or 2.5·105 (saddle-point/tetra-pod) localizations/second.


2017 ◽  
Author(s):  
Yiming Li ◽  
Markus Mund ◽  
Philipp Hoess ◽  
Ulf Matti ◽  
Bianca Nijmeijer ◽  
...  

AbstractWe present a fitter for 3D single-molecule localization of arbitrary, experimental point spread functions (PSFs) that reaches minimum uncertainty for EMCCD and sCMOS cameras, and achieves more than 105 fits/s. We provide tools to robustly model experimental PSFs and correct for depth induced aberrations, which allowed us to achieve an unprecedented 3D resolution with engineered astigmatic PSFs, and acquire high quality 3D superresolution images even on standard microscopes without 3D optics.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yongtao Liu ◽  
Zhiguang Zhou ◽  
Fan Wang ◽  
Günter Kewes ◽  
Shihui Wen ◽  
...  

AbstractSub-diffraction limited localization of fluorescent emitters is a key goal of microscopy imaging. Here, we report that single upconversion nanoparticles, containing multiple emission centres with random orientations, can generate a series of unique, bright and position-sensitive patterns in the spatial domain when placed on top of a mirror. Supported by our numerical simulation, we attribute this effect to the sum of each single emitter’s interference with its own mirror image. As a result, this configuration generates a series of sophisticated far-field point spread functions (PSFs), e.g. in Gaussian, doughnut and archery target shapes, strongly dependent on the phase difference between the emitter and its image. In this way, the axial locations of nanoparticles are transferred into far-field patterns. We demonstrate a real-time distance sensing technology with a localization accuracy of 2.8 nm, according to the atomic force microscope (AFM) characterization values, smaller than 1/350 of the excitation wavelength.


Sign in / Sign up

Export Citation Format

Share Document