中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (34): 5492-5496.doi: 10.3969/j.issn.2095-4344.1414

• 生物材料基础实验 basic experiments of biomaterials • 上一篇    下一篇

三种小鼠眼球组织石蜡切片固定液固定效果的比较

孙  良1，徐  林2
  

  1. 山东省立第三医院，1重症医学科，2中心实验室，山东省济南市  250000
  • 收稿日期:2019-05-09 出版日期:2019-12-08 发布日期:2019-12-08
  • 通讯作者: 徐林，工程师，山东省立第三医院中心实验室，山东济南市 250000
  • 作者简介:孙良，男，1981年生，山东省莱西市人，汉族，主治医师，主要从事重症感染、组织病理学研究。

Comparison of the fixation effects of three kinds of fixatives on mouse eyeball tissue

Sun Liang1, Xu Lin2
  

  1. 1Department of Critical Care Medicine, 2Central Laboratory, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China
  • Received:2019-05-09 Online:2019-12-08 Published:2019-12-08
  • Contact: Xu Lin, Engineer, Central Laboratory, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China
  • About author:Sun Liang, Attending physician, Department of Critical Care Medicine, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China

PDF

147

摘要：

文章快速阅读：

 

文题释义：
石蜡切片：组织学常规制片技术中最为广泛应用的方法，石蜡切片不仅用于观察正常细胞组织的形态结构，也是病理学和法医学等学科用以研究、观察及判断细胞组织的形态变化的主要方法，而且也已被相当广泛地用于其他许多学科领域的研究中。教学中，光镜下观察切片标本多数是石蜡切片法制备的。活的细胞或组织多为无色透明，各种组织间和细胞内各种结构之间均缺乏反差，在一般光镜下不易清楚区别出；组织离开机体后很快就会死亡和产生组织腐败，失去原有正常结构，因此组织要经固定、石蜡包埋、切片及染色等步骤以免细胞组织死亡，而能清晰辨认其形态结构。
 
 
背景：固定液是影响病理制片质量的重要因素，为了优化眼球组织病理学观察的条件，需要相对简便、实用和可靠的固定液。
目的：比较3种固定液对小鼠眼球组织石蜡切片的影响，优化小鼠眼球组织石蜡切片的固定方法。
方法：取30只BALB/c小鼠(济南朋悦实验动物繁育有限公司提供)眼球，随机分为3组，每组10只，分别固定于3种固定液中，即甲醛-戊二醛混合固定液、Davidson固定液和体积分数10%甲醛固定液，24 h后制作标本并进行苏木精-伊红染色，分析比较固定效果。动物实验获得山东省立第三医院伦理委员会批准，批准号：sdsldsyy-m-201801006。
结果与结论：①各组大部分眼球大体标本均略有收缩变形；②苏木精-伊红染色显示，甲醛-戊二醛组眼球壁出现皱缩，晶状体结构较完整、清晰，中央区无破碎，细胞排列整齐有序；角膜成纤维细胞间出现较大空隙；视网膜组织明显脱落分离，各层细胞不清晰，细胞间可见明显空白裂隙；③苏木精-伊红染色显示，Davidson组视网膜轻微变薄，分层排列较平整，未见明显缺失，无明显脱落，各层细胞排列紧密整齐，细胞核各层核膜清晰，细胞无收缩、膨胀；角膜各层结构完整，无明显结构改变，未见脱片现象，各层结构清晰、无裂隙；晶状体结构中央区可见不同程度破碎、脱片、染色不连续；④苏木精-伊红染色显示，甲醛组角膜组织轻微收缩，角膜上皮各层结构较整齐、层次较分明，晶状体和视网膜结构基本完整，但晶状体组织皱缩破裂、空隙形成，细胞排列乱序，可见视网膜组织脱落，各层排列较平整，细胞结构基本清晰，细胞间可见明显空白裂隙；⑤结果表明对于苏木精-伊红染色，Davidson固定液对角膜和视网膜的固定效果较为理想，而甲醛-戊二酸混合固定液是固定小鼠眼球晶状体组织的较好方法。

关键词: 小鼠眼球, 石蜡切片, 苏木精-伊红染色, 甲醛-戊二醛混合固定液, Davidson固定液, 10%甲醛溶液, 角膜, 晶状体

Abstract:

BACKGROUND: Fixative is a major factor affecting the quality of pathological sections. To optimize the conditions of pathological observation of eyeball tissue, a relatively simple, practical, and reliable fixative is required.
OBJECTIVE: To compare the effects of three fixative solutions on paraffin sections of mouse eyeball tissues and optimize the fixation methods.
METHODS: Thirty eyeballs of BALB/c mice (Jinan Pengyue Experimental Animal Breeding Co., Ltd., China) were randomly divided into three groups (n = 10 eyeballs/group). The harvested eyeballs were fixed in three kinds of fixatives separately, namely formaldehyde-glutaraldehyde fixative (group A), Davidson’s fixative (group B) and 10% formaldehyde fixative (group C). Twenty-four hours later, specimens were prepared for hematoxylin-eosin staining, and the fixation effects were compared. Animal experiments were approved by the Animal Ethics Committee, Shandong Provincial Third Hospital, China (approval No. sdsldsyy-m-201801006).
RESULTS AND CONCLUSION: The eyeballs of each group were all slightly contracted and deformed. Hematoxlyin-eosin staining revealed that in the group A, the eyeball wall was shrunk; the tissue structure of the lens was relatively complete and clear without fragments in the center, and cells arranged well; large gaps appeared between corneal fibroblasts; retinal tissue was clearly detached and separated, with cells in each layer unclear; obvious blank fissures were visible between cells. In the group B, the retinal was slightly thinned, the layered arrangement was flat, with no obvious loss or shedding; the cells in each layer were arranged neatly, the nuclear membrane of each layer of the nucleus was clear, the cells did not shrink or expand; the layers of the cornea were intact and had no obvious structural changes, with no peeling phenomenon; the structure of each corneal layer was clear with no crack; different degrees of fragmentation and peeling were observed in the center of the lens, with discontinuous staining. In the group C, the corneal tissue was slightly contracted, the layers of the corneal epithelium were relatively neat, the layers were relatively clear; the lens and retinal structure were basically intact, but the lens tissue shrank and ruptured; vacant space formed, cells were arranged in disorder, retinal tissue fell off, the layers were arranged evenly; cell structure was basically clear, and obvious blank fissures were visible between cells. These results suggest that for hematoxylin-eosin staining, Davidson’s fixative is ideal for the fixation of the cornea and retina and formaldehyde-glutaraldehyde fixative is better for fixing mouse eyeball lens tissue.   

Key words: mouse eyeball, paraffin section, hematoxylin-eosin staining, formaldehyde-glutaraldehyde fixative, Davidson’s fixative, 10% formaldehyde, cornea, lens

中图分类号: