Abstract

A prominent challenge in single-molecule localization microscopy is the real-time, fast, and accurate localization of nano-objects moving in three-dimensional (3D) samples. A well-established method for 3D single-molecule localization is the double-helix pointspread-function (DH-PSF) engineering, which uses additional optical elements to make the PSF exhibit different rotation angles with different nanoparticle depths. However, the compact main lobe size, effective detection depth, and precise conversion between rotation angle and depth are necessary, posing challenges to the DH-PSF generation method. Here we generate a more compact DH-PSF using Fresnel-zone-based spiral phases, and the pure phase mask achieves high transmission efficiency. The final generated DH-PSFs have a linear rotation rate at each axial position, showing a more accurate rotation angle and depth conversion. The Cramer–Rao lower limit calculation results show that the axial depth of DH-PSF extends to ${\sim}{11}\;\unicode{x00B5}{\rm m}$ with an axial localization precision of ${\sim}{45}\;{\rm nm}$ at 3000 photons and average background noise of 15. We measured the diffusion coefficient of nanospheres in different concentrations of glycerol using the generated DH-PSF. The measured results are within 6% error from the theoretical values, indicating the superior performance of the DH-PSF for nanoparticle diffusion coefficient measurements.

© 2021 Optical Society of America

Full Article  |  PDF Article
More Like This
Single shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function

Zhaojun Wang, Yanan Cai, Yansheng Liang, Xing Zhou, Shaohui Yan, Dan Dan, Piero R. Bianco, Ming Lei, and Baoli Yao
Biomed. Opt. Express 8(12) 5493-5506 (2017)

Nanoscale three-dimensional single particle tracking by light-sheet-based double-helix point spread function microscopy

Bin Yu, Jie Yu, Weihai Li, Bo Cao, Heng Li, Danni Chen, and Hanben Niu
Appl. Opt. 55(3) 449-453 (2016)

Three-dimensional parallel particle manipulation and tracking by integrating holographic optical tweezers and engineered point spread functions

Donald B. Conkey, Rahul P. Trivedi, Sri Rama Prasanna Pavani, Ivan I. Smalyukh, and Rafael Piestun
Opt. Express 19(5) 3835-3842 (2011)

References

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Data Availability

All the data in this paper are presented in the body of the text.

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Figures (7)

You do not have subscription access to this journal. Figure files are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Equations (9)

You do not have subscription access to this journal. Equations are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Metrics