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A spherical aberration-free microscopy system for live brain imaging.

Yoshihiro Ue1, Hiromu Monai2, Kaori Higuchi1

  • 1BSI-Olympus Collaboration Center, RIKEN, Hirosawa, Wako-City, 351-0198 Saitama, Japan; OLYMPUS Corporation, Hachioji-City, 192-0033 Tokyo, Japan.

Biochemical and Biophysical Research Communications
|April 13, 2018
PubMed
Summary

We developed an automated system, Deep-C, to correct spherical aberration (SA) during in vivo mouse brain imaging. This system enables clearer, quantitative analysis of neural structures deep within the brain.

Keywords:
Cerebral cortexDendritic spineRefractive indexSpherical aberrationTwo-photon microscopy

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Area of Science:

  • Neuroscience
  • Biomedical Optics
  • Microscopy

Background:

  • High-resolution in vivo imaging of mouse brains requires high numerical aperture (NA) and long working distance (WD) objectives.
  • Spherical aberration (SA) from refractive index mismatch hinders imaging deep in the brain, worsening with depth.
  • Existing manual SA correction methods cause focal shifts, complicating precise imaging.

Purpose of the Study:

  • To develop an automated system for spherical aberration (SA) compensation during in vivo brain imaging.
  • To enable stable, high-resolution, and quantitative analysis of fine neural structures at depth.
  • To overcome limitations of manual SA correction and focal drift.

Main Methods:

  • An objective-attached device was created to coordinate collar rotation and Z-position for automated SA correction.
  • A fast iterative algorithm was developed to determine optimal SA correction based on image contrast.
  • The system, named Deep-C, was tested with a 25x water-immersion objective for in vivo two-photon microscopy.

Main Results:

  • The Deep-C system effectively reduced SA in mouse cerebral cortical layers, enabling quantitative analysis of YFP-labeled neurons.
  • The degree of SA compensation allowed for assessment of refractive indices in different cortical layers.
  • Fully corrected 3D reconstructions of neurons in live mouse brains were achieved.

Conclusions:

  • The automated Deep-C system successfully compensates for SA in heterogeneous tissues like the mouse brain.
  • This technology enhances the performance of correction-collar-equipped objectives for deep tissue imaging.
  • Deep-C facilitates reproducible, quantitative analysis and 3D reconstruction of neural circuits in vivo.