大鼠尾椎间盘不同切片方式及染色方法的组织学观察比较

doi:10.12300/j.issn.1674-5817.2020.108

摘要/Abstract

摘要： 目的比较大鼠尾椎间盘不同切片方式及染色方法的优缺点,为相应的临床诊断及实验研究奠定理论基础。方法收集SD大鼠尾椎间盘,制成冷冻切片及石蜡切片后,分别行苏木精-伊红染色（HE染色）、番红-固绿染色、Masson染色、甲苯胺蓝染色,以及针对一型胶原蛋白（typeⅠcollagen,COL-Ⅰ）、二型胶原蛋白（typeⅡcollagen,COL-Ⅱ）和糖胺聚糖（glycosaminoglycan,GAG）的免疫荧光染色,对比观察椎间盘各层组织形态及蛋白表达的情况。结果在染色方法中,石蜡切片HE染色能清晰显示各层组织;Masson染色能清晰显示纤维环组织及髓核组织;番红-固绿染色及甲苯胺蓝染色能较好地显示软骨各层及软骨下骨结构。冷冻切片的免疫荧光染色中,纤维环组织的胶原蛋白多为COL-Ⅰ,髓核组织中胶原蛋白多为COL-Ⅱ,而GAG多由髓核细胞及髓核样细胞分泌。结论观察椎间盘各层组织时,选用HE染色及Masson染色等染色,其中石蜡切片优于冷冻切片;观察椎间盘各种蛋白的表达及分布时,选用免疫荧光染色,其中冷冻切片优于石蜡切片。

关键词: 椎间盘, 石蜡切片, 冷冻切片, 免疫荧光, 特殊染色, 大鼠

Abstract: Objective To compare the advantages and disadvantages of different slicing and staining methods of caudal intervertebral disc in rats, so as to provide a theoretical basis for clinical diagnosis and experimental research. Methods The caudal intervertebral discs of SD rats were collected and made into frozen sections and paraffin sections. Hematoxylin-eosin staining (HE staining), safranin O-fast green staining, Masson staining, and toluidine blue staining of the caudal intervertebral disc sections, and immunofluorescence staining of typeⅠ collagen (COL-Ⅰ), typeⅡcollagen (COL-Ⅱ) and glycosaminoglycan (GAG) were performed to observe the tissue morphology and protein expression. Results Of the selected staining methods, HE staining of the paraffin sections clearly showed each layer structure of intervertebral discs; Masson staining results showed fibrous annulus tissues and nucleus pulposus tissues; safranin O-fast green staining and toluidine blue staining results showed cartilage layers and subchondral bone structures. In the frozen sections with immunofluorescence staining, COL-Ⅰ was dominant in fibrous annulus, COL-Ⅱwas dominant in nucleus pulposus, and GAG was mainly secreted by nucleus pulposus cells and nucleus pulposus-like cells. Conclusion When observing the layer structure of intervertebral discs, staining techniques such as HE staining and MASSON staining should be chosen, and paraffin sections are better than frozen sections. When observing the expression and distribution of various proteins of intervertebral discs, immunofluorescence should be selected, and frozen sections are superior to paraffin sections.

Key words: Intervertebral disc, Paraffin section, Frozen section, Immunofluorescence, Special staining, Rats

中图分类号:

R-33
Q95-33

赵宇朋, 周平辉, 官建中. 大鼠尾椎间盘不同切片方式及染色方法的组织学观察比较[J]. 实验动物与比较医学, 2021, 41(2): 100-107.

ZHAO Yupeng, ZHOU Pinghui, GUAN Jianzhong. Comparison of Different Slicing and Staining Methods of Caudal Intervertebral Discs in Rats[J]. Laboratory Animal and Comparative Medicine, 2021, 41(2): 100-107.

图/表 4

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