Micropatterning assembly, methods for micropatterning, and micropatterned devices

A method for producing a system comprising a cell binding device for modular micropatterning one or more cell binding molecules comprising:

• a) providing a solid carrier comprising a surface;
• b) passivating the surface of the solid carrier by covalently attaching a polymeric coating to the surface of the solid carrier;
• c) contacting the passivated surface of the solid carrier with a liquid phase comprising one or more adaptor molecules, wherein each of the one or more adaptor molecules comprises:
• (i) a terminal photoreactive moiety configured to covalently photoimmobilize the adaptor molecule to a predetermined micropatterning area of the passivated polymeric coating on the solid carrier surface by photoinduced radical formation, wherein the terminal photoreactive moiety is a fluorescent dye, benzophenone, or phenylazide, and
• (ii) a terminal click chemistry cycloaddition moiety that is chemically distinct from and orthogonal to the photoreactive moiety, wherein the terminal click chemistry cycloaddition moiety is an alkyne, cyclic alkyne, transcycloocetene, norbornene, cyclopropane, or tetrazine, and the terminal click chemistry cycloaddition moiety is configured to covalently bind a counterpart reactive group present on the one or more cell binding molecules by cycloaddition subsequent to photoimmobilization of the adaptor molecule by the terminal photoreactive moiety,
• wherein the photoreactive moiety and the click chemistry cycloaddition moiety are covalently attached and spatially separated;
• d) projecting a desired light pattern onto the solid carrier surface at the liquid interface to selectively activate the photoreactive moiety, thereby covalently immobilizing the adaptor molecules to predetermined micropatterning areas of the passivated polymeric coating while maintaining the click chemistry cycloaddition moiety in a chemically unaltered and available state for subsequent covalent binding;
• and
• e) contacting the one or more immobilized adaptor molecules with the one or more cell binding molecules under conditions that enable the click chemistry cycloaddition moiety to covalently bind with the counterpart reactive group on the cell binding molecules by cycloaddition, thereby producing a micropatterned cell binding device.