《中国康复理论与实践》

《中国康复理论与实践》

• 基础研究 • 上一篇    下一篇

两种组织透明技术在免疫荧光染色观察脊髓3D结构中的应用①

段红梅 1，尚俊奎 2，郝鹏 2，李千千 1，杨飞祥 2，杨朝阳 1,2，李晓光 1,2   

  1. 1.北京航空航天大学生物与医学工程学院，北京市 100191；2.首都医科大学神经生物学系，北京市 100069。
  • 出版日期:2016-04-25 发布日期:2016-06-01

Application of Two Kinds of Tissue Clearing Method in Observing 3D Imaging of Spinal Cord with Immunofluorescent Staining

DUAN Hong- mei1, SHANG Jun- kui2, HAO Peng2, LI Qian- qian1, YANG Fei- xiang2, YANG Zhao- yang1,2, LI Xiao-guang1,2   

  1. 1.Department of Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, China; 2.Department of Neurobiology, Capital Medical University, Beijing 100069, China
  • Published:2016-04-25 Online:2016-06-01

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摘要： 目的 观察 CUBIC和 iDISCO两种透明技术在脊髓免疫荧光染色中的效果。方法 应用两种组织透明技术处理 1 mm厚脊髓冠状切片，然后行神经丝(NF)蛋白免疫荧光染色，激光共聚焦扫描显微镜观察染色效果。结果 与CUBIC透明相比，iDISCO透明技术所需时间短，透明度高(F=6.64, P<0.01)，免疫荧光染色深(F=5117.55, P<0.01)。结论 应用iDISCO透明方法和免疫荧光染色相结合的方法可以完整观察脊髓轴突，更加简单快速，染色效果更好。

关键词: 脊髓, 透明, 轴突再生, CUBIC, iDISCO, 大鼠

Abstract: Objective To compare the application of CUBIC and iDISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and iDISCO, respectively. The neurofilament (NF) protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, iDISCO had the advantages of shorter time, higher transparency (F=6.64, P<0.01), and deeper penetration (F= 5117.55, P<0.01). Conclusion Immunofluorescent staining combined with iDISCO could completely observe the spinal axons with shorter time and better stain effect.

Key words: spinal cord, transparency, axon regeneration, CUBIC, iDISCO, rats