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Quantification of light attenuation in optically cleared mouse brains.

Angela d'Esposito1, Daniil Nikitichev2, Adrien Desjardins2

  • 1University College London, Centre for Advanced Biomedical Imaging, 72 Huntley Street, London WC1E6DD, United Kingdom.

Journal of Biomedical Optics
|August 18, 2015
PubMed
Summary

Optical clearing techniques like BABB and pBABB effectively clear biological tissues for 3D imaging. This study quantifies clearing efficacy, finding lipid-rich regions attenuate light the most.

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Area of Science:

  • Biomedical Engineering
  • Optical Imaging
  • Tissue Clearing

Background:

  • Optical clearing techniques enhance deep tissue imaging by reducing light scattering and absorption.
  • Quantifying the efficacy of different tissue clearing methods is crucial for optimizing imaging protocols.

Purpose of the Study:

  • To develop and apply a spectroscopy-based method for quantifying optical clearing efficacy in biological tissues.
  • To compare the clearing performance of BABB, pBABB, and passive CLARITY methods in mouse brains.

Main Methods:

  • Developed a custom spectroscopy system to measure light attenuation in cleared tissues.
  • Applied the system to quantify clearing quality in mouse brains subjected to three different clearing protocols.
  • Analyzed regional light attenuation in relation to tissue composition, specifically lipid content.

Main Results:

  • BABB (benzyl alcohol and benzyl benzoate) and pBABB (peroxide BABB) demonstrated the highest degree of optical clearing.
  • The developed method allows for regional measurements of light attenuation.
  • Higher lipid content in specific brain regions correlated with increased light attenuation.

Conclusions:

  • The developed spectroscopy approach provides a quantitative measure of optical clearing efficacy.
  • BABB and pBABB are effective clearing agents for mouse brain tissue.
  • Understanding regional light attenuation is important for interpreting imaging data from cleared tissues and selecting appropriate clearing protocols.