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Maximizing the field of view and accuracy in 3D Single Molecule Localization Microscopy.

Published version
Peer-reviewed

Change log

Authors

Rehman, Sohaib Abdul 
Carr, Alexander R 
Lenz, Martin O 
Lee, Steven F 

Abstract

Super-resolution techniques that localize single molecules in three dimensions through point spread function (PSF) engineering are very sensitive to aberrations and optical alignment. Here we show how double-helix point spread function is affected by such mis-alignment and aberration. Specifically, we demonstrate through simulation and experiment how misplacement of phase masks in infinity corrected systems is a common source of significant loss of accuracy. We also describe an optimal alignment and calibration procedure to correct for these errors. In combination, these optimizations allow for a maximal field of view with high accuracy and precision. Though discussed with reference to double-helix point spread function (DHPSF), the optimization techniques are equally applicable to other engineered PSFs.

Description

Keywords

0299 Other Physical Sciences, Bioengineering

Journal Title

Opt Express

Conference Name

Journal ISSN

1094-4087
1094-4087

Volume Title

26

Publisher

The Optical Society
Sponsorship
Medical Research Council (MR/K015850/1)
Engineering and Physical Sciences Research Council (EP/M003663/1)
Engineering and Physical Sciences Research Council (EP/L015455/1)