Comparison of Different Slicing and Staining Methods of Caudal Intervertebral Discs in Rats

doi:10.12300/j.issn.1674-5817.2020.108

Abstract

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

CLC Number:

R-33
Q95-33

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.

Figures/Tables 4

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