中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (6): 1348-1356.doi: 10.4103/1673-5374.329473

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

组织光学透明技术能快速、精确和全面评估神经组织三维形态

  

  • 出版日期:2022-06-15 发布日期:2021-12-17

Optical tissue clearing enables rapid, precise and comprehensive assessment of three-dimensional morphology in experimental nerve regeneration research

Simeon C. Daeschler1, *, Jennifer Zhang1, 3, Tessa Gordon1, 3, Gregory H. Borschel1, 2, 3, 4   

  1. 1SickKids Research Institute, Neuroscience and Mental Health Program, Toronto, ON, Canada; 2Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada; 3Division of Plastic and Reconstructive Surgery, the Hospital for Sick Children, Toronto, ON, Canada; 4Indiana University School of Medicine, Indianapolis, IN, USA
  • Online:2022-06-15 Published:2021-12-17
  • Contact: Simeon C. Daeschler, MD, simeondaeschler@gmail.com.
  • Supported by:
    This work was supported by the German Research Foundation (DA 2255/1-1, to SCD).

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摘要：

既往研究对于神经组织的形态学分析主要局限于组织切片。新的组织光学透明技术能够对整个器官进行三维成像。为了扩大其在实验性神经修复研究中的适用性，实验将这些技术用于大鼠的神经及其靶器官。组织光学透明技术与传统的实验室技术相匹配，包括逆行标记研究和计算图像分割，可进行快速和精确的细胞定量分析。此外，组织光学透明技术处理的器官能够以前所未有的规模进行三维形态测量，包括全皮节神经支配研究、追踪肌肉内神经分支或绘制神经血管网络图。鉴于其广泛的适用性、快捷性和低廉的成本，组织光学透明技术可能成为下一代神经修复研究的关键技术。

https://orcid.org/0000-0001-8130-7714 (Simeon C. Daeschler)

Abstract: Morphological analyses are key outcome assessments for nerve regeneration studies but are historically limited to tissue sections. Novel optical tissue clearing techniques enabling three-dimensional imaging of entire organs at a subcellular resolution have revolutionized morphological studies of the brain. To extend their applicability to experimental nerve repair studies we adapted these techniques to nerves and their motor and sensory targets in rats. The solvent-based protocols rendered harvested peripheral nerves and their target organs transparent within 24 hours while preserving tissue architecture and fluorescence. The optical clearing was compatible with conventional laboratory techniques, including retrograde labeling studies, and computational image segmentation, providing fast and precise cell quantitation. Further, optically cleared organs enabled three-dimensional morphometry at an unprecedented scale including dermatome-wide innervation studies, tracing of intramuscular nerve branches or mapping of neurovascular networks. Given their wide-ranging applicability, rapid processing times, and low costs, tissue clearing techniques are likely to be a key technology for next-generation nerve repair studies. All procedures were approved by the Hospital for Sick Children’s Laboratory Animal Services Committee (49871/9) on November 9, 2019.

Key words: histology, image segmentation, morphometry, nerve regeneration, outcome assessment, peripheral nerve, three-dimensional imaging, tissue clearing

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