连续切片三维重构技术在黑腹果蝇中肠超微结构研究中的应用

Application of the serial section 3D reconstruction technique to the study of the midgut ultrastructure of Drosophila melanogaster

【目的】黑腹果蝇Drosophila melanogaster中肠超微结构的观察对于其功能的研究具有重要意义。本研究利用扫描电镜及三维重构技术,旨在探究黑腹果蝇中肠超微结构。【方法】解剖野生型黑腹果蝇并取其中肠,经四氧化锇-硫代甲酰二肼-四氧化锇(OTO)方法制备中肠样本后,进行连续切片,并通过扫描电镜对连续切片上的中肠组织超微结构进行观察,利用Amira软件对中肠特定结构进行三维重构。【结果】黑腹果蝇中肠超微结构包括浆膜腔屏障、肌层、围食膜、粘液层、肠道微生物、微气管、肠道上皮细胞、肠道干细胞和肠分泌细胞;肠上皮细胞包含细胞核、线粒体、微绒毛和迷路结构。通过三维重构展示了肠道微绒毛,线粒体,肠腔内容物,微气管和迷路。【结论】三维重构能更加精确全面展示中肠超微结构。

[Objectives]To investigate the ultrastructure of the midgut of Drosophila melanogaster using scanning electron microscopy and three-dimensional reconstruction,thereby improving understanding of its function.[Methods]Midguts were removed from wild-type D.melanogaster,continuously sectioned and prepared for scanning electron microscopy using the osmium tetroxide-thiocarbohydrazide-osmium tetroxide(OTO)method.The ultrastructure of midgut tissue was then observed under a scanning electron microscope.Specific structures of the midgut were reconstructed in 3D with Amira software.[Results]Midgut ultrastructure consisted of a serosal barrier,muscle layer,perinatal membrane,mucous layer,intestinal microorganisms,tracheole,intestinal epithelial cells,intestinal stem cells and enteroendocrine cells.Intestinal epithelial cells contain mitochondria,microvilli and basal labyrinths.A three-dimensional reconstruction of the intestinal microvilli,mitochondria,luminal contents,microtrachea and basal labyrinths,was created.[Conclusion]3D reconstruction was an effective way to accurately and comprehensively visualize the ultrastructure of the D.melanogaster midgut.