Study on immunohistochemical staining techniques of human inner ear frozen sections

doi:10.3969/j.issn.1006-7795.2022.06.014

Journal of Capital Medical University ›› 2022, Vol. 43 ›› Issue (6): 905-910.doi: 10.3969/j.issn.1006-7795.2022.06.014

• New Progress of Rhinology Research • Previous Articles Next Articles

Study on immunohistochemical staining techniques of human inner ear frozen sections

Li Ying^1,2, Chen Biao¹, Wang Naili³, Hao Xinping^1*

1. Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;
2. Beijing Institute of Otolaryngology, Key Laboratory of Otolaryngology Head and Neck Surgery (Capital Medical University), Ministry of Education, Beijing Key Laboratory of Nasal Diseases, Beijing 100005, China;
3. Morphological Laboratory, Teaching Center, Institute of Basic Medicine, Chinese Academy of Medical Sciences, Beijing 100005,China

Received:2022-09-06 Online:2022-12-21 Published:2022-11-30
Contact: ^*E-mail:13671218615@139.com

Abstract

Abstract: Objective To study the methods of anti-slice escaping treatment and antigenic retrieval in the process of immunohistochemical staining of frozen sections of human inner ear for an improved staining. Methods Temporal bone specimens without ear diseases were taken. After sampling, fixation, decalcification, dehydration, and embedding, frozen sections were performed and frozen at -20 ℃. According to the anti-slice escaping treatment, the sample was divided into three groups: post-fixation group (group I), baked slice group (group Ⅱ), and post-fixation combined with baked slice group (group Ⅲ). Then each group underwent antigen retrieval and was divided into group A (heat repair group), group B (enzymatic digestion group) and group C (Triton group).The expression of four antibodies of WARP, NF200, CD68 and IBA1 in each group was detected by indirect fluorescence immunohistochemistry, observed under laser confocal microscope, and the results of dissection were analyzed. Results Compared to group Ⅰ and group Ⅱ, group Ⅲ had the best effect on preventing slices escape from glass slid. The antigen retrieval effect of group A and group C was relatively satisfactory. While the repair effect of group B was not good with a destructive effect on the cellular structure of the tissue. Conclusion The application of post-fixation combined with baked slice method to anti-slice escaping and the application of heat repair or Triton treatment for antigen repair have achieved good results in this study. Appropriate anti-slice escaping treatment and antigen retrieval methods play a crucial role in improving the sensitivity of immunohistochemical staining and preparation quality of inner ear frozen sections.

Key words: human inner ear, frozen section, anti-slice escaping treatment, antigen retrieval, indirect immunofluorescence technique

CLC Number:

R764.3

Li Ying, Chen Biao, Wang Naili, Hao Xinping. Study on immunohistochemical staining techniques of human inner ear frozen sections[J]. Journal of Capital Medical University, 2022, 43(6): 905-910.

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Study on immunohistochemical staining techniques of human inner ear frozen sections

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