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Analysis of Organelle Positioning Using Patterned Microdevices

机译:用图案化微型器材分析细胞器定位

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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 howorganelle 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.
机译:在许多疾病中观察到细胞内细胞器分布的改变的后果往往尚不清楚。细胞内细胞器改变它们的形态和定位以调节细胞稳态和功能。我们概述了如何研究Howorganelle定位,采用基于密度的3D图像分析,其应用于显示类似蜂窝几何形状的细胞。通过使用在微图案化基材上播种的单细胞来促进定量,该细胞提供用于控制电池扩散的提示。这种最小的系统模拟了通常在组织中观察到的细胞器的可再现分布,简化图像分析并最小化观察鲁棒表型所需的细胞数。在这里,我们提供了大多数细胞器可以在粘合微图案上播种的细胞中有效分析大多数细胞器的指导方针。我们举例说明了由于细胞骨架或相关电动机蛋白的扰动而导致不同细胞器的定位的改变是有效量化的。

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