陈思怀 李德龙 高继业

摘要 为了克服小鼠组织器官传统石蜡切片制作方法存在组织易碎、结构不完整的缺点，对传统石蜡切片制作进行了改良。将浸蜡前的透明步骤改为异硬脂醇与石蜡混合液处理，替代苯类透明剂制作空肠、脾脏2种器官的石蜡切片，并以切片机切片时蜡带质量和HE染色后镜下质量为切片制作效果的评定指标。结果表明，2种器官在切片机上均能切出完整、较长的蜡带，组织块无硬化、不碎裂，展片平整;HE染色过程中不掉片，染色后光镜下细胞质与细胞核染色对比清晰，器官组织结构完整。就脾脏和空肠而言，应用异硬脂醇与石蜡混合液替代苯类透明剂能制作出优良的石蜡切片。

关键词 石蜡切片;异硬脂醇;透明剂;小鼠

中图分类号 R-361.2  文献标识码 A

文章编号 0517-6611（2021）11-0094-02

doi：10.3969/j.issn.0517-6611.2021.11.025

開放科学（资源服务）标识码（OSID）：

Improvement of Preparation Method of Paraffin Sections of Some Tissues and Organs in Mice

CHEN Si-huai，LI De-long，GAO Ji-ye

（College of Veterinary Medicine，Southwest University， Chongqing 402460）

Abstract In order to overcome the defects of fragile tissue and incomplete structure in the traditional paraffin section preparation method of mice tissues and organs， the traditional preparation method of paraffin section was improved.Paraffin sections about jejunum and spleen were made with the mixed solution of isostearin and paraffin instead of benzene transparent agents.The quality of ribbon and microscopic observation after HE staining were selected as judgment indices .The results showed that two kinds of representative organs could be cut into long and complete wax bandes on the microtome， and flatten out with no hardening or non-fragmentation.After HE staining， the cytoplasm and nucleus were stained clearly， and the structures of organs and tissues were intact.As for jejunum and spleen，the mixture of isostearic alcohol and paraffin could be used to treat the tissues instead of benzene transparent agents to make good paraffin sections.

Key words Paraffin section;Isostearic alcohol;Transparent agent;Mice

小鼠作为实验动物，其组织器官在大小、质地等方面不同于成年畜禽，如小肠壁薄、肝细胞脂类丰富、脾脏小等。在开展教学或病理石蜡切片制作时，采用传统制片方法，以二甲苯作为浸蜡前透明剂，容易造成切片破碎、组织分离或结构龟裂。鉴于二甲苯等苯类物质对健康的危害以及处理组织块需特定技术与经验等缺点，许多学者报道了应用无毒的硬脂酸（十八烷酸）与石蜡的混合液替代浸蜡前二甲苯透明剂的使用，并成功制作出优良的动物组织石蜡切片[1-2]。但是，该方法有以下缺点：一是细胞内硬脂酸若未被石蜡置换完全，容易导致染色掉片，影响镜下观察效果;二是硬脂酸熔点高，与石蜡混合需较高的温度（≥67 ℃）来保持熔融状态，这使小鼠组织器官更容易硬化、脆化[3]。……