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Three-Photon Adaptive Optics for Mouse Brain Imaging.

David Sinefeld1,2, Fei Xia1,3, Mengran Wang1

  • 1School of Applied and Engineering Physics, Cornell University, Ithaca, NY, United States.

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|June 13, 2022
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Summary
This summary is machine-generated.

Three-photon microscopy (3PM) with adaptive optics (AO) enables deeper, clearer imaging of mouse brains. This advanced technique improves resolution and signal quality in scattering tissues, even at depths over 1 mm.

Keywords:
adaptive opticsbrain imagingin vivo imagingmultiphoton microcopythree-photon microscopy

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Area of Science:

  • Neuroscience
  • Biomedical Optics
  • Microscopy

Background:

  • Three-photon microscopy (3PM) offers deeper tissue penetration than two-photon microscopy (2PM) due to longer wavelengths and reduced scattering.
  • Adaptive optics (AO) techniques, utilizing spatial light modulators (SLMs), can correct optical aberrations to enhance imaging depth and resolution.
  • In vivo imaging of scattering biological tissues like the mouse brain presents significant challenges for microscopy.

Purpose of the Study:

  • To present and analyze a three-photon microscopy adaptive optics (3PM AO) system for enhanced in vivo mouse brain imaging.
  • To demonstrate the capability of 3PM AO for deep-tissue imaging and aberration correction in scattering brain environments.
  • To characterize the performance and limitations of AO correction within the mouse brain using 3PM.

Main Methods:

  • Development and implementation of a 3PM AO system using a 1300 nm femtosecond laser source.
  • Utilized a microelectromechanical (MEMS) SLM for wavefront aberration correction via Zernike phase patterns.
  • Employed three-photon (3P) fluorescence signal as feedback for aberration correction, avoiding direct phase measurement.

Main Results:

  • Achieved significant signal improvement in the cortex and hippocampus at depths exceeding 1 mm.
  • Demonstrated near-diffraction-limited imaging in cortical layers, including visualization of dendritic spines.
  • Characterized the effective volume for AO correction in mouse brain tissue.

Conclusions:

  • The developed 3PM AO system effectively enhances imaging depth and resolution in scattering mouse brain tissue.
  • The study validates the use of 3P fluorescence feedback for AO correction in in vivo microscopy.
  • Understanding the limitations of AO correction is crucial for optimizing deep-brain imaging applications.