组织透明化三维显示全包埋耳皮肤神经血管网  

Tissue Clearing for Three-dimensional Visualization of The Neurovascular Network in The Whole Mount Auricular Skin

作　　者：张锴文 郭亚婷 苏雨昕 王雨晴 崔晶晶[1] 张建梁[1] 王佳[1] 白万柱[1] ZHANG Kai-Wen;GUO Ya-Ting;SU Yu-Xin;WANG Yu-Qing;CUI Jing-Jing;ZHANG Jian-Liang;WANG Jia;BAI Wan-Zhu(Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院针灸研究所,北京100700

出　　处：《生物化学与生物物理进展》2023年第6期1473-1479,共7页Progress In Biochemistry and Biophysics

基　　金：中国中医科学院科技创新工程(CI2021A03407);国家自然科学基金(82004492);中国中医科学院优秀青年科技人才(创新类)培养专项(ZZ14-YQ-032)资助项目。

摘　　要：目的结合组织学染色技术和组织透明化策略研究耳部皮肤中神经纤维和血管的空间对应关系。方法将耳廓前面和后面的皮肤从其中间软骨仔细剥离,然后直接分别用蛋白基因产物9.5(PGP 9.5)和鬼笔环肽对耳部皮肤的神经纤维和血管进行免疫荧光染色并进行组织透明化处理。随后,以全包埋方式将耳皮肤组织裱贴在载玻片上用于荧光显微镜和共聚焦显微镜观察。结果本研究显示PGP 9.5阳性神经纤维伴随鬼笔环肽标记的血管一道从耳廓的基部走行到其外围区域形成耳廓的神经血管网。在传统的免疫荧光染色技术基础上,后续的组织透明化技术可以更好地展示耳部皮肤中神经纤维和血管的形态学细节。结论从方法学的角度来看,组织透明化技术增进了耳部皮肤中免疫荧光标记的可视性,它可能成为有效的技术手段用于解析正常和病理状态下耳部神经血管网的空间结构。Objective To study the spatial correlation of the nerve fibers and blood vessels in the auricular skin by using the combination of histological staining technique and tissue clearing strategy.Methods The anterior and posterior auricular skins were carefully peeled away from the intervening cartilage and the auricular nerve fibers and blood vessels were directly immunofluorescent stained with protein gene product 9.5(PGP 9.5)and phalloidin followed with tissue clearing treatment.After that,these auricular skins were mounted on the microscope slide in the whole mount pattern and examined under an epifluorescence microscope and a laser scanning confocal microscope.Results It was shown that the PGP 9.5+nerve fibers aligning with phalloidin-labeled blood vessels ran from the base part of auricle to its peripheral region,forming the neurovascular network in the auricular skin.Beyond the conventional immunofluorescence staining,the additional tissue clearing treatment demonstrated the auricular nerve fibers and blood vessels better in morphological detail.Conclusion From the perspective of methodology,tissue clearing technique enhance the visualization of the immunofluorescence labeling within the auricular skin,which should be an effective approach for insight into the auricular neurovascular network in a three-dimensional view under the physiological and pathological conditions.

关 键 词：组织透明化 耳部皮肤 神经纤维 血管 神经血管网 

分 类 号：G312[文化科学]