scholarly journals High-order-helix point spread functions for monocular three-dimensional imaging with superior aberration robustness

2018 ◽  
Vol 26 (4) ◽  
pp. 4873 ◽  
Author(s):  
René Berlich ◽  
Sjoerd Stallinga
Keyword(s):  
Three Dimensional ◽  
High Order ◽  
Point Spread Functions ◽  
Three Dimensional Imaging ◽  
Point Spread

scholarly journals Analyzing engineered point spread functions using phasor-based single-molecule localization microscopy

2020 ◽  
Author(s):  
Koen J.A. Martens ◽  
Abbas Jabermoradi ◽  
Suyeon Yang ◽  
Johannes Hohlbein
Keyword(s):  
Saddle Point ◽  
Single Molecule ◽  
Software Package ◽  
Double Helix ◽  
Three Dimensional ◽  
Fourier Plane ◽  
Point Spread Functions ◽  
Localization Microscopy ◽  
Point Spread ◽  
Single Molecule Localization Microscopy

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.

Sign in / Sign up

Export Citation Format

Share Document