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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
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Deep Volumetric Super-Resolution Imaging in Thick Biological Specimens With Sparse Scanning SIM.

Sha An1,2,3, Xuhong Guo1,2,3, Zhongxia Cai1,2,3

  • 1School of Physics, Xidian University, Xi'an, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|February 23, 2026
PubMed
Summary
This summary is machine-generated.

Sparse scanning SIM (SS-SIM) enables deep, super-resolution imaging in thick biological samples. This advanced microscopy technique overcomes scattering limitations for clearer, volumetric visualization of dense tissues and organisms.

Keywords:
deep super‐resolution imagingresonant scanningstructured illumination microscopy

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Area of Science:

  • Biophotonics and advanced microscopy techniques.
  • Development of novel imaging modalities for biological research.

Background:

  • High-resolution fluorescence microscopy faces challenges in deep, thick specimens due to light scattering and limited penetration.
  • Structured illumination microscopy (SIM) offers super-resolution but is sensitive to scattering, hindering volumetric imaging.
  • Existing methods struggle with deep imaging in complex biological environments.

Purpose of the Study:

  • To develop a super-resolution microscopy technique for deep imaging in thick, live specimens.
  • To overcome the limitations of conventional SIM in scattering-prone biological samples.
  • To enable high-contrast, volumetric visualization of dense biological structures.

Main Methods:

  • Introduction of sparse scanning SIM (SS-SIM), integrating rapid laser scanning with pixel-addressed intensity modulation and sCMOS detection.
  • Utilization of either one- or two-photon excitation for versatile deep imaging applications.
  • Implementation with water-immersion objectives to achieve imaging depths up to 300/600 µm.

Main Results:

  • SS-SIM achieved a spatial resolution of approximately 150 nm.
  • Maintained a 1.6/1.7-fold (lateral/axial) resolution enhancement over wide-field microscopy within the imaging depth.
  • Successfully enabled volumetric visualization of dense biological samples, including thick tissues, organoids, and small organisms.

Conclusions:

  • SS-SIM provides a robust and simple method for deep super-resolution imaging.
  • This technique significantly expands the applicability of SIM to challenging biological samples.
  • SS-SIM offers a practical solution for high-contrast volumetric imaging in complex biological environments.