Rab11/FIP5复合体的三维延时分析：顶端管腔形成过程中的时空动态变化
3D time-lapse analysis of Rab11/FIP5 complex: spatiotemporal dynamics during apical lumen formation.
作者信息
Mangan Anthony, Prekeris Rytis
机构信息
Department of Cell and Developmental Biology, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Mail Stop 8108, 12801 East 17th Avenue, Aurora, CO, 80045, USA.
出版信息
Methods Mol Biol. 2015;1298:181-6. doi: 10.1007/978-1-4939-2569-8_15.
Fluorescent imaging of fixed cells grown in two-dimensional (2D) cultures is one of the most widely used techniques for observing protein localization and distribution within cells. Although this technique can also be applied to polarized epithelial cells that form three-dimensional (3D) cysts when grown in a Matrigel matrix suspension, there are still significant limitations in imaging cells fixed at a particular point in time. Here, we describe the use of 3D time-lapse imaging of live cells to observe the dynamics of apical membrane initiation site (AMIS) formation and lumen expansion in polarized epithelial cells.
对在二维（2D）培养物中生长的固定细胞进行荧光成像，是观察蛋白质在细胞内定位和分布最广泛使用的技术之一。尽管该技术也可应用于在基质胶基质悬浮液中生长时形成三维（3D）囊肿的极化上皮细胞，但在对特定时间点固定的细胞进行成像时仍存在重大限制。在此，我们描述了使用活细胞的三维延时成像来观察极化上皮细胞中顶端膜起始位点（AMIS）形成和管腔扩张的动态过程。
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