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Adaptive optics improves multiphoton super-resolution imaging.

Wei Zheng1,2, Yicong Wu1, Peter Winter1

  • 1Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.

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Summary

We enhanced multiphoton microscopy resolution and intensity using a nonlinear guide star and deformable mirror. This technique improves deep tissue imaging in various biological samples.

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Biophysics

Background:

  • Multiphoton microscopy offers deep tissue imaging but suffers from optical aberrations.
  • Structured illumination microscopy enhances resolution but is sensitive to aberrations.

Purpose of the Study:

  • To improve resolution and signal intensity in multiphoton structured illumination microscopy.
  • To correct optical aberrations in deep biological tissues.

Main Methods:

  • Utilized a nonlinear guide star to measure optical aberrations.
  • Employed a deformable mirror for real-time aberration correction.
  • Applied the technique to various biological samples including cells, larvae, and Drosophila brains.

Main Results:

  • Achieved up to 40-fold increase in peak intensity.
  • Recovered resolution to 176 ± 10 nm laterally and 729 ± 39 nm axially.
  • Demonstrated effective aberration correction at depths of ~250 micrometers.

Conclusions:

  • The developed method significantly enhances the performance of multiphoton structured illumination microscopy.
  • This technique enables clearer and deeper imaging of biological structures.
  • Offers a valuable tool for advanced biological research and diagnostics.