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

作者:Rehman Sohaib Abdul*; Carr Alexander R; Lenz Martin O; Lee Steven F; O'Holleran Kevin
来源:Optics Express, 2018, 26(4): 4631-4637.
DOI:10.1364/OE.26.004631

摘要

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. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License.

  • 出版日期2018-2-19