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
王 博；黄风景；王林燕；罗 军；关 旸
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DOI：10.7679/j.issn.2095-1353.2022.085
作者单位：浙江中医药大学中医药科学院，杭州 310053；温州医科大学附属第二医院中医科，温州 325027； 浙江丰虹新材料股份有限公司，湖州 313300
中文关键词：黑腹果蝇；中肠；超微结构；扫描电镜；三维重构
英文关键词：Drosophila melanogaster; midgut; ultrastructure; SEM; 3D reconstruction
中文摘要：
【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
英文摘要：
[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.