García-Arcos Juan M, Gateau Kevin, Venkova Larisa, Piel Matthieu
Institut Pierre Gilles de Gennes, PSL Research University, Paris, France.
Current affiliation, Department of Biochemistry and Swiss National Centre of Competence in Research, Chemical Biology, University of Geneva, Geneva, Switzerland.
Methods Mol Biol. 2023;2608:63-81. doi: 10.1007/978-1-0716-2887-4_5.
Physical confinement in microfluidic devices has become a common technique to induce and study cell migration in a large range of cell types. Confined migration was previously understudied due to the limitations of 2D migration assays but has emerged as an important mode of migration in the past decade. Furthermore, confinement improves the quality of the imaging and simplifies the analysis of trajectories by confining migration to the plane of acquisition. Protocols described in this chapter relate to methods extending the previously published 2D confinement technique. First, we explain a method to increase the complexity of the confinement chamber by microfabricating nanometer-sized PDMS grooves on the bottom surface, usually used for contact guidance studies. Then, we describe a method to perform the confinement on cells embedded inside a μm-thin 3D collagen gel. Finally, we describe an alternative method to confine cells based on agarose, so that cells can be fixed or drug perfused while being confined, which is currently not possible in the 2D confinement silicone-based device.
在微流控设备中进行物理限制已成为诱导和研究多种细胞类型细胞迁移的常用技术。由于二维迁移分析的局限性，受限迁移此前研究较少，但在过去十年中已成为一种重要的迁移模式。此外，通过将迁移限制在采集平面内，限制提高了成像质量并简化了轨迹分析。本章所述方案涉及扩展先前发表的二维限制技术的方法。首先，我们解释一种通过在底面微制造纳米尺寸的聚二甲基硅氧烷（PDMS）凹槽来增加限制室复杂性的方法，该方法通常用于接触导向研究。然后，我们描述一种对嵌入微米级薄三维胶原蛋白凝胶内的细胞进行限制的方法。最后，我们描述一种基于琼脂糖限制细胞的替代方法，以便在限制细胞的同时对其进行固定或药物灌注，而这在基于硅酮的二维限制设备中目前是无法实现的。
