Capmany Anahi, Latgé Bruno, Schauer Kristine
Institut Curie, PSL Research University, Molecular Mechanisms of Intracellular Transport Group, Paris, France.
Centre National de la Recherche Scientifique, Paris, France.
Curr Protoc Cell Biol. 2019 Mar;82(1):e77. doi: 10.1002/cpcb.77. Epub 2018 Nov 15.
The consequences of alterations in the distribution of intracellular organelles, observed in many diseases, are often not clear. Intracellular organelles alter their morphology and positioning to regulate cell homeostasis and function. We outline how organelle positioning can be studied employing a density-based analysis of 3D images applied to cells that show similar cellular geometries. Quantification is facilitated by the use of single cells seeded on micropatterned substrates that provide cues for controlled cell spreading. This minimal system mimics the reproducible distribution of organelles typically observed in tissues, simplifying image analysis and minimizing the number of cells required for the observation of robust phenotypes. Here we provide guidelines for how the majority of organelles can be efficiently analyzed in cells seeded on adhesive micropatterns. We exemplify how alterations in the positioning of different organelles as a result of the perturbation of the cytoskeleton or associated motor proteins can be efficiently quantified. © 2018 by John Wiley & Sons, Inc.
在许多疾病中观察到的细胞内细胞器分布改变的后果往往并不清楚。细胞内细胞器会改变其形态和定位以调节细胞内稳态和功能。我们概述了如何通过对具有相似细胞几何形状的细胞应用基于密度的三维图像分析来研究细胞器定位。使用接种在微图案化底物上的单细胞有助于定量分析，这些底物可为细胞的可控铺展提供线索。这个最小化系统模拟了组织中通常观察到的细胞器可重复分布，简化了图像分析并减少了观察到稳定表型所需的细胞数量。在这里，我们提供了如何在接种于粘性微图案上的细胞中有效分析大多数细胞器的指南。我们举例说明了由于细胞骨架或相关运动蛋白的扰动导致不同细胞器定位改变时如何进行有效定量分析。© 2018约翰威立国际出版公司
