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Imaging and positioning through scattering media with double-helix point spread function engineering.

Jingjing Gao1,2, Pengwei Wang1, Wenwen Li3

  • 1Chinese Academy of Sciences, Shanghai Institute of Optics and Fine Mechanics, Key Laboratory of Quantum Optics, Shanghai, China.

Journal of Biomedical Optics
|April 28, 2023
PubMed
Summary
This summary is machine-generated.

This study demonstrates super-resolution microscopy through scattering media using modified double-helix point spread function (DH-PSF) microscopy. The technique achieves high 3D localization accuracy, enabling clearer imaging of biological samples within turbid environments.

Keywords:
deconvolutiondouble-helix point spread functionpenetration thicknessscattering mediumthree-dimensional localization

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Double-helix point spread function (DH-PSF) microscopy enables 3D super-resolution imaging but is limited to non-scattering environments.
  • Super-resolution imaging through turbid or scattering media remains a significant challenge in microscopy.

Purpose of the Study:

  • To investigate the feasibility of DH-PSF microscopy for imaging and localization within scattering environments.
  • To enhance 3D localization accuracy and imaging quality in turbid media.

Main Methods:

  • Modification of the conventional DH-PSF method with a scanning strategy and deconvolution algorithm.
  • Localization of fluorescent microspheres by identifying the center of double spots.
  • Image reconstruction via deconvolution of the DH-PSF from scanned data.

Main Results:

  • Achieved localization accuracy of 13 nm (transverse) and 51 nm (axial).
  • Demonstrated imaging penetration up to an optical thickness (OT) of 5.
  • Presented proof-of-concept 3D localization and imaging through biological membranes (eggshell, onion epidermis).

Conclusions:

  • Modified DH-PSF microscopy successfully images and localizes targets in scattering media with super-resolution.
  • The method offers a potential solution for deeper and clearer visualization in scattering environments.
  • Enables in situ super-resolution microscopy for demanding applications using various fluorescent probes.