Photopatterning with a printed transparency mask and a protein-friendly photoresist

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.