豚鼠耳蜗在不同中间液中扫描电镜制样效果的比较研究  

Effects of Different Intermediate Preparation Media on Scanning Electron Microscopy Images of Guinea Pig Cochleae

作　　者：曹鋐 崔颖 CAO Hong;CUI Ying(Department of Otorhinolaryngology Head and Neck Surgery,First Affiliated Hospital of Jinzhou Medical University)

机构地区：[1]锦州医科大学附属第一医院耳鼻咽喉头颈外科

出　　处：《中华耳科学杂志》2021年第4期626-629,共4页Chinese Journal of Otology

摘　　要：目的用无水乙醇代替醋酸异戊酯作为临界点干燥过程中的中间液,提高扫描电镜制样过程中效率,减少在实验过程中醋酸异戊酯挥发可能造成伤害。方法6只2月龄听力正常豚鼠,1%戊巴比妥钠腹腔注射麻醉。2.5%戊二醛灌流双侧耳蜗,进行解剖,后固定,脱水等流程进行至临界点干燥法。无水乙醇和醋酸异戊酯分别作为中间液进行临界点干燥。将干燥后的耳蜗进行喷金处理,于扫描电镜下进行观察。对比不同中间液对耳蜗不同位置毛细胞纤毛,表皮板等结构的影响。结果无水乙醇作为中间液制备的扫描电镜样本减少了用醋酸异戊酯作为中间液的步骤,并且耳蜗毛细胞纤毛及表皮板等超微结构保存完好。两种中间液制备的样本在扫描电镜观察下效果均良好。结论在豚鼠耳蜗样本制备中无水乙醇能够代替醋酸异戊酯作为中间液在临界点干燥法中的应用,能够确保耳蜗超微结构的完整性,缩短实验时间,减少了醋酸异戊酯因挥发对空气环境的污染,保护了实验人员的安全。Objective Using anhydrous ethanol in place of isoamyl acetate in the intermediate medium during critical point drying,to improve the efficiency of sample preparation for scanning electron microscopy with shortened time and reduced hazards from isoamyl acetate vapor during preparation.Methods Six guinea pigs with normal hearing and aged 2 months were anesthetized by intraperitoneal injection of 1%pentobarbital sodium.Both cochlea were perfused with 2.5%glutaraldehyde under an anatomical microscope,and then harvested,fixed,dehydrated and prepared for conventional scanning electron microscopy until critical point drying.Anhydrous ethanol or isoamyl acetate was used as the intermediate solution for critical point drying.The dried cochlea was sprayed with gold and examined under a scanning electron microscope.Images of the hair cell cilium and epidermal plate at different locations in cochlear samples prepared with anhydrous ethanol or isoamyl acetate were compared.Results Processing steps were reduced when using anhydrous ethanol as intermediate medium with ultrastructures of hair cell cilia and epidermal plate well preserved and comparable to when using isoamyl acetate.Conclusion In guinea pigs,,the application of anhydrous ethanol can replace isoamyl acetate as the intermediate liquid during critical point drying in cochlea sample preparation,which can ensure the integrity of cochlear ultrastructure,shorten preparation time,reduce air pollution by volatile isoamyl acetate and improve safety of the laboratory personnel.

关 键 词：临界点干燥法 耳蜗 醋酸异戊酯 无水乙醇 

分 类 号：R764[医药卫生—耳鼻咽喉科]