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Related Experiment Video

Updated: Jul 17, 2026

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

Efficient double helix detection with steerable filters.

Andrew E S Barentine1, Ashwin Balaji1,2, W E Moerner1,3

  • 1Department of Chemistry, Stanford University, Stanford, California 94305, USA.

Biomedical Optics Express
|July 16, 2026
PubMed
Summary

We developed an efficient method for localizing double helix point-spread functions in 3D microscopy. This technique uses steerable filters for fast and accurate 3D single-molecule localization and tracking.

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Efficient Double Helix Detection with Steerable Filters.

bioRxiv : the preprint server for biology·2025

Area of Science:

  • Biophysics
  • Microscopy
  • Computational Biology

Background:

  • 3D single-molecule localization microscopy (SMLM) is crucial for visualizing cellular structures.
  • Accurate localization of molecules is essential for high-resolution 3D imaging.
  • Current methods can be computationally intensive and require significant user input.

Purpose of the Study:

  • To develop an efficient and user-friendly detection scheme for localizing double helix point-spread functions (PSFs) in 3D SMLM.
  • To reduce computational cost and complexity compared to existing deep learning approaches.
  • To integrate the scheme into an open-source platform for broader accessibility.

Main Methods:

  • Utilized steerable filters for extracting 2D position and lobe orientation (axial position) estimates.

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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
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A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging
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A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

Published on: April 8, 2019

Related Experiment Videos

Last Updated: Jul 17, 2026

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging
08:13

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

Published on: April 8, 2019

  • Implemented a detection and fitting pipeline with percentile-based background subtraction and signal-to-noise-based thresholding.
  • Developed a plug-in for the open-source Python Microscopy Environment (PYME) for parallel analysis.

    • Main Results:

    • Achieved efficient localization of double helix PSFs using only 7 convolutions.
    • Demonstrated a significant reduction in computational complexity compared to deep learning methods.
    • The complete pipeline demonstrated state-of-the-art performance with minimal user intervention.

    Conclusions:

    • The presented detection scheme offers an efficient and accurate method for 3D SMLM.
    • The integration into PYME enhances accessibility and usability for researchers.
    • This approach provides a powerful tool for advancing single-molecule localization and tracking studies.