大鼠和小鼠阴道组织神经血管层的全层铺片技术探索  

Exploration of full-thickness slice technique of the neurovascular layer of vaginal tissue

作　　者：于普光 冯家劲 占学康 李健 任帅 陈科明 韦秋绫 何瑞 李广永[3] YU Puguang;FENG Jiajin;ZHAN Xuekang;LI Jian;REN Shuai;CHEN Keming;WEI Qiuling;HE Rui;LI Guangyong(College of Clinical Medicine,Ningxia Medical University,Yinchaun 750004,China;Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education,Ningxia Medical University;Department of Urology,General Hospital of Ningxia Medical University)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]宁夏医科大学生育力保持教育部重点实验室 [3]宁夏医科大学总医院泌尿外科

出　　处：《山西医科大学学报》2022年第9期1191-1194,共4页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81860268)。

摘　　要：目的通过不同实验条件建立一种稳定、可重复的用于显示阴道神经与血管的全层铺片技术。方法雌性SD大鼠和ICR小鼠各20只,麻醉后分离出阴道组织,分别用中性缓冲福尔马林和丙酮固定,每种固定液分别固定24 h和48 h后,在显微镜下逐层剥离出阴道肌层制作铺片,观察不同固定方式制作铺片的脱片率,并行HE染色、Masson染色、嗜银染色和免疫组化观察组织的形态、肌纤维、血管和神经组织。结果同种固定液固定24 h和48 h在制作铺片的难易程度上无明显差别。不同的固定液和不同的固定时间在是否容易脱片方面无显著差异(Fisher确切概率法,P_(小鼠)=0.893和P_(大鼠)=0.429)。中性缓冲福尔马林固定后组织较韧,更易剥离各层组织,而丙酮固定的组织易碎,组织不完整。不同的固定方法对HE染色、Masson染色和嗜银染色结果影响不大,但对免疫组化染色会产生较大影响,即中性缓冲福尔马林固定使抗体更容易结合。结论两种固定液都能很好地对阴道组织起固定作用,中性缓冲福尔马林固定液用于制作阴道肌层全层铺片优于丙酮,该铺片技术能很好地显示阴道组织中的肌肉、神经与血管组织,可用于今后的研究。Objective To establish a repeatable and stable full-thickness technique for displaying vaginal nerves and blood vessels under various experimental conditions.Methods After anesthesia,the vaginal tissues of 20 female SD rats and 20 female ICR mice were removed and then fixed with neutral buffered formalin and acetone,respectively.The vaginal muscle layer was removed to make slides,and the detachment rate of slides created using various fixation methods was measured.HE staining,Masson staining,silver staining,and immunohistochemistry were used to observe the morphology of tissues,blood vessels,and nerve tissues.Results There was no significant difference in the difficulty of making slides with the same fixative for 24 h or 48 h.There was no statistically significant difference in decapsulation between two fixative solutions for different fixation time(Fisher’s exact test,P_(ICR)=0.893 and P_(SD)=0.429).The tissue was tougher and easier to drop after neutral buffer formalin fixation,whereas acetone-fixed tissue was fragile and incomplete.Different fixation methods had little effect on the results of HE staining,Masson staining,and argentophilic staining,but had a greater impact on immunohistochemical staining,namely,the neutral buffered formalin fixation made antibodies bind more easily.Conclusion Both fixatives are effective to fix vaginal tissue.The neutral buffered formalin fixative has better efficacy to create a full-thickness vaginal muscle layer than acetone.This technique can better display muscle,nerve,and vascular tissue,which is suitable for the further research.

关 键 词：阴道 神经血管层 全层铺片技术 

分 类 号：R446[医药卫生—诊断学]