3种组织固定液对大菱鲆肝脏切片质量影响和效果比较  

Effect Comparison of Three Tissue Fixation Solutions on Liver Section Quality of Turbot(Scophthalmus maximus)

作　　者：秦宏宇 高云涛 高云红 李明月 贾玉东[1,2] QIN Hongyu;GAO Yuntao;GAO Yunhong;LI Mingyue;JIA Yudong(Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao,Shandong 266071;Qingdao National Laboratory for Marine Science and Technology,Qingdao,Shandong 266237;Tianjin Agricultural University Fisheries College,Tianjin 300384)

机构地区：[1]中国水产科学研究院黄海水产研究所,山东青岛266071 [2]青岛海洋科学与技术试点国家实验室,山东青岛266237 [3]天津农学院水产学院,天津300384

出　　处：《中国农学通报》2023年第5期148-153,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目“IGF系统关键基因调控大菱鲆卵母细胞减数分裂阻滞与恢复机制研究”(31972811);中国水产科学研究院基本科研业务费项目“陆海接力养殖创新团队”(2020TD51)。

摘　　要：为查明3种组织固定液对大菱鲆肝脏石蜡切片质量的影响,筛选效果最佳的固定液。本实验选用波恩氏(Bouin)、戴维森氏(Davidson)和4%多聚甲醛(Paraformaldehyde, PFA)3种组织固定液,对大菱鲆的肝脏进行固定,制作石蜡切片,通过苏木素-伊红(H-E)和免疫组织化学(Immunohistochemistry)染色,观察组织细胞形态学特征,利用统计学方法分析细胞完整度和免疫组化阳性率情况。结果发现,3种固定液对大菱鲆肝组织石蜡切片固定效果存在显著差异,Bouin固定液组织H-E染色细胞结构清晰、形态完整,Davidson固定液H-E染色组织溶解、细胞结构不清晰,但能分辨出核质界线,4%多聚甲醛组H-E染色鲜艳,但核质分界不清晰。完整肝细胞比例,4%PFA显著低于Bouin和Davidson固定液(P<0.05),且Bouin和Davidson固定液无显著差异(P>0.05)。肝组织增殖细胞核抗原(proliefrating cell nuclear antigen, PCNA)免疫组化染色阳性率3种固定液存在显著差异,Bouin固定液显著高于4%PFA和Davidson(P<0.05)。综上,使用Bouin固定液对肝脏固定,进行H-E染色或免疫组化染色能观察到更为完整的组织细胞结构和细胞增殖情况,效果最佳。In order to identify the influence of three fixation solutions on the quality of paraffin section of turbot liver tissue, and select the best fixation solution, in this study, three fixation solutions, Bouin fixation solution,Davidson fixation solution and 4% Paraformaldehyde(PFA), were used to fix the turbot liver tissue, and paraffin sections were prepared. The morphological characteristics of the tissue cells were observed by hematoxylin-eosin(H-E) and immunohistochemical staining. The integrity and immunohistochemical positive rate of hepatocyte were analyzed by statistical methods. The results showed that there were significant differences in the fixation effect of the three fixation solutions on paraffin sections of turbot liver tissue. Clear structure and complete morphology of hepatocyte were observed after H-E staining with Bouin fixation solution. However, the hepatocyte manifested dissolved and unclear structure by Davidson fixation solution, but the nucleocytoplasmic boundary could be distinguished. 4% PFA fixation solution showed bright color of hepatocyte, but the boundary of nucleus and plasma was not clear. The percentage of intact hepatocytes after4% PFA fixation was significantly lower than that after Bouin and Davidson fixation solutions treatment(P<0.05), Bouin and Davidson fixation solutions showed no significant difference(P>0.05). There were significant differences in the positive rate of hepatic proliefrating cell nuclear antigen(PCNA) immunohistochemical staining among the three fixation solutions, and that of Bouin fixation solution was significantly higher than 4%PFA and Davidson fixation solutions(P<0.05). In conclusion, Bouin fixation solution is the optimal selection for turbot hepatic H-E or immunohistochemical staining for observing tissue cell structure and cell proliferation.

关 键 词：组织固定液 苏木素-伊红染色 免疫组织化学 增殖细胞核抗原 肝脏 

分 类 号：S966[农业科学—水产养殖]