Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Oct 7;21(10):1873–1883. doi: 10.1038/s41592-024-02382-8

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 5 — a, The bias on the estimation of the out-of-plane angle θ as a function of rotational mobility γ for the intensity-only algorithm, determined using simulated images of single dipole emitters (using 1,000 photons per emitter and ten background photons per pixel). A rotational mobility parameter γ of 0 corresponds to total rotational freedom, and a value of 1 corresponds to perfect rotational immobilization. b, The same as a but for the DSF-fitting algorithm. c, Lateral localization precision as a function of the number of detected photons as determined from simulations (ten background photons per pixel, γ = 1). Separate curves are shown for molecules at different out-of-plane orientations. d, The same as c but for the precision of ϕ estimation. e, The computer processing time for the datasets from f for the intensity (int.)-only algorithm with centroid localization, the intensity-only algorithm with least-squares fitting of a rotated (rot.) asymmetric Gaussian (Gauss) and the DSF-fitting algorithm. Refer to the Methods for computer specifications. f, A POLCAM SMOLM reconstruction of a 200 × 200-pixel region from Fig. 4 with 10,000 frames, as analyzed by the intensity-only algorithm (with rotated asymmetric Gaussian fitting) and the DSF-fitting algorithm. Both reconstructions were rendered using approximately the same number of localizations (258,251 localizations for the intensity-only algorithm, 258,172 localizations for the DSF-fitting algorithm). g, Insets from the regions in f that are marked by dotted boxes. The insets display some structural differences between the reconstructions generated by the two algorithms.