Heuzé Mélina L, Collin Olivier, Terriac Emmanuel, Lennon-Duménil Ana-Maria, Piel Matthieu
Subcellular Structure and Cellular Dynamics, Institut Curie, Paris, Cedex, France.
Methods Mol Biol. 2011;769:415-34. doi: 10.1007/978-1-61779-207-6_28.
This chapter describes a method to study cells migrating in micro-channels, a confining environment of well-defined geometry. This assay is a complement to more complex 3D migration systems and provides several advantages even if it does not recapitulate the full complexity of 3D migration. Important parameters such as degree of adhesion, degree of confinement, mechanical properties, and geometry can be varied independently of each other. The device is fully compatible with almost any type of light microscopy and the simple geometry makes automated analysis very easy to perform, which allows screening strategy. The chapters is divided into five parts describing the design of different types of migration chambers, the fabrication of a mold by photolithography, the assembly of the chamber, the loading of cells, and finally the imaging on live or fixed cells.
本章介绍了一种研究细胞在微通道（一种几何形状明确的受限环境）中迁移的方法。该检测方法是对更复杂的三维迁移系统的补充，即使它不能完全重现三维迁移的全部复杂性，但仍具有几个优点。诸如粘附程度、受限程度、机械性能和几何形状等重要参数可以相互独立地变化。该装置与几乎任何类型的光学显微镜完全兼容，其简单的几何形状使自动分析非常容易进行，这有利于筛选策略。本章分为五个部分，分别描述了不同类型迁移腔室的设计、通过光刻制造模具、腔室的组装、细胞加载，以及最后对活细胞或固定细胞的成像。
