摘要
Abstract
Ultrathin sectioning technology has been widely used as an important means of veterinary research.However,the general preparation procedure of ultrathin sectioning technology cannot obtain high-quality results for different samples.Based on the traditional electron microscopy technology,this study first optimized the ultrathin sectioning technology of sheep skins,and then used the optimized method to verify the application in characterizing DTMUV infected duck brain tissues and BHK-21 cells,microsporidia infected Portunus trituberculatus muscle tissues and PRV infected Vero cells.The optimized procedure included 4％ paraformaldehyde modified fixative,acetone as dehydrating agent in the whole process,30％ dehydration step,100％ dehydration by time gradients for 10 min,15 min,20 min and 30 min,double immersion and a pure embedding agent.Therefore,the optimized steps stably obtained flat and uniform slices.The use of 20％ethanol configuration of uranium acetate solution and extension of the staining time to 20 minutes made the slices clean,pollution-free and staining contrast.The 75-mesh large aperture mesh greatly increased the observed area of the slices.These optimizations enabled to process high-quality ultrathin sections with large field of view as visualized in DTMUV-infected duck brain tissues and BHK-21 cells,microsporidia-infected muscle tissues of Portunus trituberculatus and PRV-infected Vero cells.These results confirmed that the optimized ultrathin sectioning technique met the needs of veterinary ultrastructural morphology research.关键词
超薄切片/透射电镜/皮肤/鸭坦布苏病毒/微孢子虫/伪狂犬病毒Key words
Ultrathin slice/transmission electron microscope/skin/DTMUV/microsporidia/PRV分类
农业科技
