不同组织固定液对毛囊内蛋白固定效果的对比研究  

Comparative study on the fixation effect of different tissue fixatives on proteins in hair follicles

作　　者：袁燕平 汪亮亮 胡佳莉 Yuan Yanping;Wang Liangliang;Hu Jiali(Department of Pathology,Jiujiang University Affiliated Hospital,Jiujiang 332000)

机构地区：[1]九江学院附属医院病理科,江西九江332000

出　　处：《中国现代医药杂志》2024年第4期11-15,共5页Modern Medicine Journal of China

基　　金：江西省卫生健康委科技计划项目(编号:2023131077)。

摘　　要：目的探讨4%多聚甲醛、10%中性缓冲甲醛液、丙酮和95%乙醇4种组织固定液对大鼠毛囊冷冻切片免疫组织荧光染色的影响,找出最佳的冰冻切片固定液。方法选取2只SD大鼠,乙醚麻醉后处死,剪下大鼠唇部皮肤,分离触须毛囊后随即进行冷冻连续切片。将切片分别置于4%多聚甲醛、10%中性缓冲甲醛液、丙酮、95%乙醇固定液中固定,所有切片经过相同的固定时间后进行免疫荧光染色,检测毛囊内细胞角蛋白(CK15)和β-连环蛋白(β-catenin)的表达情况。结果免疫荧光染色结果显示,4%多聚甲醛、10%中性缓冲甲醛、丙酮、95%乙醇对蛋白抗原的保存效果各不相同,其中4%多聚甲醛固定效果较好,组织内可见较强的荧光表达,而经10%中性缓冲甲醛、丙酮、95%乙醇固定的组织则呈现更低的表达状态。结论不同的组织固定液对蛋白的保存效果存在差异,4%多聚甲醛对毛囊内蛋白具有更好的保存效果。Objective To investigate the effects of 4% paraformaldehyde,10% neutral buffered formaldehyde solution,and 95% ethanol as four types of tissue fixatives on immunofluorescence staining of frozen sections of rat hair follicles,and identify the optimal fixative for frozen sections.Methods Two SD rats were randomly selected and anesthetized with ether.The skin of the rat lips was cut off,and the tentacle hair follicles were isolated and immediately performed serial frozen sections.The sections were fixed in four different fixatives including 4% paraformaldehyde,10% neutral buffered formaldehyde solution,acetone and 95% ethanol.After the same fixation time,all sections were subjected to immunofluorescence staining to detect the expression of CK15 and β-catenin proteins in the hair follicles.Results The immunofluorescence staining results showed that the preservation effect of four different fixatives on proteins were different.Among them,4% paraformaldehyde had better preservation effect,and strong fluorescence expression was observed in the tissues.However,tissues fixed with 10% neutral buffered formaldehyde solution,acetone and 95%ethanol showed lower expression levels.Conclusion There are differences in the preservation effect of different tissue fixatives on proteins,and 4% paraformaldehyde has better preservation effect on proteins in hair follicles.

关 键 词：冰冻切片 固定液 4%多聚甲醛 免疫荧光 蛋白 

分 类 号：R361.2[医药卫生—病理学]