Hao Marlene M, Bergner Annette J, Newgreen Donald F, Enomoto Hideki, Young Heather M
Department of Anatomy and Neuroscience, University of Melbourne, Melbourne, VIC, Australia.
Laboratory for Enteric Neuroscience, TARGID, University of Leuven, Leuven, Belgium.
Methods Mol Biol. 2019;1976:97-105. doi: 10.1007/978-1-4939-9412-0_8.
Time-lapse imaging of gut explants from embryonic mice in which neural crest-derived cells express fluorescent proteins allows the behavior of enteric neural crest cells to be observed and analyzed. Explants of embryonic gut are dissected, mounted on filter paper supports so the gut retains its tubular three-dimensional structure, and then placed in coverglass bottom culture dishes in tissue culture medium. A stainless steel ring is placed on top of the filter support to prevent movement. Imaging is performed using a confocal microscope in an environmental chamber. A z series of images through the network of fluorescent cells is collected every 3, 5, or 10 min. At the end of imaging, the z series are projected.
对来自胚胎小鼠的肠道外植体进行延时成像，其中神经嵴衍生细胞表达荧光蛋白，这使得肠道神经嵴细胞的行为能够被观察和分析。解剖胚胎肠道外植体，将其置于滤纸支架上，以便肠道保持其管状三维结构，然后将其置于盖玻片底部的培养皿中，加入组织培养基。在滤纸支架顶部放置一个不锈钢环以防止移动。使用共聚焦显微镜在环境箱中进行成像。每隔3、5或10分钟收集一系列通过荧光细胞网络的z轴图像。成像结束时，将z轴系列图像进行投影。
