Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell.
Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell.
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DOI：
10.3791/52918
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发表时间：
2015-07-19
期刊：
影响因子：
--
通讯作者：
Liddle RA
中科院分区：
文献类型：
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作者：
Bohórquez D;Haque F;Medicetty S;Liddle RA
描绘细胞的超微结构对于理解其功能是重要的。这可能是一个令人生畏的项目，因为稀有的细胞类型遍布由不同细胞类型组成的组织，如肠上皮的肠内分泌细胞。这些食物和细菌的胃肠道传感器一直很难研究，因为它们以1：1000的比例分散在其他上皮细胞中。最近，已经产生了转基因报告小鼠，以通过荧光鉴定肠内分泌细胞。其中之一是肽YY-GFP小鼠。使用这种鼠标，我们开发了一种方法，相关的共聚焦和串行块面扫描电子显微镜。我们将该方法命名为cocem 3D，并将其应用于识别组织中特定的肠内分泌细胞，并在3D中揭示细胞的超微结构。cocem 3D的分辨率足以识别小至分泌囊泡的细胞器，并区分细胞膜的体积渲染。Cocem 3D可以很容易地适用于研究其天然组织中其他特定细胞类型的3D超微结构。
Delineation of a cell’s ultrastructure is important for understanding its function. This can be a daunting project for rare cell types diffused throughout tissues made of diverse cell types, such as enteroendocrine cells of the intestinal epithelium. These gastrointestinal sensors of food and bacteria have been difficult to study because they are dispersed among other epithelial cells at a ratio of 1:1,000. Recently, transgenic reporter mice have been generated to identify enteroendocrine cells by means of fluorescence. One of those is the peptide YY-GFP mouse. Using this mouse, we developed a method to correlate confocal and serial block-face scanning electron microscopy. We named the method cocem3D and applied it to identify a specific enteroendocrine cell in tissue and unveil the cell’s ultrastructure in 3D. The resolution of cocem3D is sufficient to identify organelles as small as secretory vesicles and to distinguish cell membranes for volume rendering. Cocem3D can be easily adapted to study the 3D ultrastructure of other specific cell types in their native tissue.