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Updated: May 27, 2025

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

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

Published on: August 27, 2014

Localizing axial dense emitters based on single-helix point spread function and compressed sensing.

Hanzhe Wu¹, Danni Chen¹, Yihong Ji¹

¹Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, Guangdong Province, China.

Nanophotonics (Berlin, Germany)

|February 20, 2025

Summary

This summary is machine-generated.

This study introduces SH-CS, a novel method for 3D single molecule localization microscopy. It enhances temporal resolution and localization accuracy for dense molecular imaging using light needle excitation and compressed sensing.

Keywords:

compressed sensing single-helix point spread function three-dimensional single molecule localization microscopy

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Area of Science:

Biophysics
Optical Microscopy
Nanotechnology

Background:

3D single molecule localization microscopy (SMLM) often sacrifices temporal resolution for localization accuracy.
Point spread function (PSF) engineering encodes depth information but typically requires sparse molecular excitation.

Purpose of the Study:

To develop a method for improving temporal resolution and axial localization accuracy in 3D SMLM.
To enable imaging of relatively dense molecular distributions.

Main Methods:

Proposed the SH-CS method combining light needle excitation, single helix-point spread function (SH-PSF) detection, and compressed sensing (CS).
Applied CS algorithm to SH images for decoding axial molecular information at each scanning position.

Main Results:

Simulations showed axial localization accuracy of 12.1-73.5 nm for 1-15 molecules within a 4 μm depth range.
Feasibility validated using a 3D sample of fluorescent beads.

Conclusions:

The SH-CS method offers improved temporal resolution and localization accuracy for 3D SMLM.
The technique is suitable for imaging relatively dense molecular samples, overcoming limitations of traditional sparse excitation methods.