Kang Jiwoo, Choi Jong-Cheol, Kim Miju, Jung Hong-Ryul, Doh Junsang
School of Interdisciplinary Bioscience and Bioengineering (I-Bio), Pohang University of Science and Technology, Pohang, Gyeongbuk, South Korea.
Department of Mechanical Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, South Korea.
Methods Cell Biol. 2014;119:55-72. doi: 10.1016/B978-0-12-416742-1.00004-4.
Microscope projection photolithography (MPP) based on a protein-friendly photoresist is a versatile tool for the fabrication of protein- and cell-micropatterned surfaces. Photomasks containing various features can be economically produced by printing features on transparency films. Features in photomasks are projected by the objective lens of a microscope, resulting in a significant reduction of the feature size to as small as ~1 μm, close to the practical limit of light-based microfabrication. A fluorescence microscope used in most biology labs can be used for the fabrication process with some modifications. Using such a microscope, multistep MPP can be readily performed with precise registration of each micropattern on transparency film masks. Here, we describe methods of the synthesis and characterization of a protein-friendly photoresist poly(2,2-dimethoxy nitrobenzyl methacrylate-r-methyl methacrylate-r-poly(ethylene glycol) methacrylate) and the setups of fluorescence microscopes and the MPP procedures. In addition, we describe the protocols used in the micropatterning of multiple lymphocytes and the dynamic micropatterning of adherent cells.
基于对蛋白质友好的光刻胶的显微镜投影光刻（MPP）是一种用于制造蛋白质和细胞微图案化表面的通用工具。通过在透明薄膜上印刷特征，可以经济地生产包含各种特征的光掩模。光掩模中的特征由显微镜的物镜投射，从而使特征尺寸显著减小至约1μm，接近基于光的微制造的实际极限。大多数生物实验室中使用的荧光显微镜经过一些修改后可用于制造过程。使用这样的显微镜，可以轻松地进行多步MPP，并在透明薄膜掩模上精确对齐每个微图案。在这里，我们描述了一种对蛋白质友好的光刻胶聚（2,2-二甲氧基硝基苄基甲基丙烯酸酯-r-甲基丙烯酸甲酯-r-聚（乙二醇）甲基丙烯酸酯）的合成和表征方法、荧光显微镜的设置以及MPP程序。此外，我们描述了在多个淋巴细胞微图案化和贴壁细胞动态微图案化中使用的方案。
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