Azioune Ammar, Carpi Nicolas, Tseng Qingzong, Théry Manuel, Piel Matthieu
Systems Cell Biology of Cell Division and Cell Polarity, UMR144, Institut Curie, CNRS, Paris 75248, France.
Methods Cell Biol. 2010;97:133-46. doi: 10.1016/S0091-679X(10)97008-8.
The described protocol is a simple method to make protein micropatterns with a micron size resolution. It can be applied to control cell shape and adhesive geometry, and also for any other assay requiring protein patterning. It is based on the use of a photomask with microfeatures to locally irradiate with deep UV light (below 200 nm) an antifouling substrate, making it locally adsorbing for proteins. The entire process can be subdivided into three main parts. The first part describes the design of a photomask. The second part describes the passivation (antifouling treatment) of the substrate, its irradiation, and the binding of proteins. The entire process can be completed in a couple of hours. It requires no expensive equipment and can be performed in any biology lab. The last part describes cell deposition on the micropatterned substrate. We also provide a discussion with pitfalls and alternative techniques adapted to various substrates, including silicone elastomers.
所述方案是一种制作具有微米级分辨率蛋白质微图案的简单方法。它可用于控制细胞形状和黏附几何形状，也适用于任何其他需要蛋白质图案化的检测。该方法基于使用具有微特征的光掩模，用深紫外光（低于200nm）对防污底物进行局部照射，使其对蛋白质具有局部吸附性。整个过程可细分为三个主要部分。第一部分描述光掩模的设计。第二部分描述底物的钝化（防污处理）、照射以及蛋白质的结合。整个过程可在几个小时内完成。它不需要昂贵的设备，可在任何生物实验室进行。最后一部分描述细胞在微图案化底物上的沉积。我们还提供了关于陷阱以及适用于各种底物（包括硅橡胶）的替代技术的讨论。
