Grossier Jean-Philippe, Goud Bruno, Schauer Kristine
Unité Mixte de Recherche, Institut Curie, Paris, France.
Methods Mol Biol. 2014;1174:117-38. doi: 10.1007/978-1-4939-0944-5_8.
Despite a large body of publications on endocytosis, only a few studies have focused on its spatial organization. To study how endocytosis is related to distinct cellular sites, we combine cell normalization by the "micropatterning technique" with the quantification of spatial organization by "probabilistic density mapping." Micropatterns of extracellular matrix proteins impose adhesive and non-adhesive areas to cultured cells and allow the control of adhesion geometry, shape, and cell organization. Probabilistic density maps provide a visual summary for 3D localization of the structures of interest and enable the extraction of robust statistics for quantification of cellular organization. Here, we provide a method to analyze and compare the spatial distribution of endocytosed ligands in micropatterned cells. This approach permits to establish the role of cellular adhesion on uptake mechanisms and to address the potential function of predefined sites of endocytosis.
尽管关于内吞作用已有大量文献发表，但只有少数研究关注其空间组织。为了研究内吞作用如何与不同的细胞位点相关，我们将通过“微图案化技术”进行的细胞标准化与通过“概率密度映射”进行的空间组织量化相结合。细胞外基质蛋白的微图案对培养细胞施加粘附和非粘附区域，并允许控制粘附几何形状、形状和细胞组织。概率密度图为感兴趣的结构的三维定位提供了直观的总结，并能够提取可靠的统计数据以量化细胞组织。在这里，我们提供了一种方法来分析和比较微图案化细胞中内吞配体的空间分布。这种方法有助于确定细胞粘附在摄取机制中的作用，并探讨内吞作用预定义位点的潜在功能。
