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Area of Science:

  • Biophysics
  • Neuroscience
  • Optical Microscopy

Background:

  • Single-molecule switching nanoscopy (SMSN) is limited by sample-induced aberrations when imaging beyond coverslips.
  • These aberrations distort and blur crucial single-molecule emission patterns, hindering deep-tissue imaging.
  • Robust 3D imaging within tissues remains a significant challenge for nanoscopy techniques.

Purpose of the Study:

  • To overcome the limitations of SMSN in scattering and aberrating biological tissues.
  • To develop a method for high-resolution 3D imaging of molecular structures within intact tissue samples.
  • To enable the visualization of nanoscale details of pathological hallmarks in neurodegenerative diseases.

Main Methods:

  • Combined active shaping of point spread functions (PSFs) with efficient adaptive optics.
  • Implemented a robust 3D-SMSN system capable of penetrating scattering media.
  • Utilized the developed system to image through 30-μm-thick mouse brain sections.

Main Results:

  • Successfully enabled robust 3D-SMSN imaging within biological tissues, overcoming sample-induced aberrations.
  • Achieved visualization and reconstruction of nanoscale details of amyloid-β filaments.
  • Demonstrated the capability to image through significant tissue depths (30 μm).

Conclusions:

  • The developed active PSF shaping and adaptive optics approach significantly enhances 3D-SMSN performance in scattering tissues.
  • This technique provides a powerful new tool for studying the nanoscale architecture of biomolecular targets in complex biological environments.
  • The successful imaging of amyloid-β filaments in an Alzheimer's disease mouse model highlights the potential for disease mechanism research.