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Micropatterned surfaces with controlled ligand tethering.

Microcontact printing (micro-CP) is a facile, cost-effective, and versatile soft-lithography technique to create two-dimensional patterns of domains with distinct functionalities that provides a robust platform to generate micropatterned biotechnological arrays and cell culture substrates. Current micro-CP approaches rely on nonspecific immobilization of biological ligands, either by direct printing or adsorption from solution, onto micropatterned domains surrounded by a nonfouling background. This technique is limited by insufficient control over ligand density. We present a modified micro-CP protocol involving stamping mixed ratios of carboxyl- and tri(ethylene glycol)-terminated alkanethiols that provides for precise covalent tethering of single or multiple ligands to prescribed micropatterns via standard peptide chemistry. Processing parameters were optimized to identify conditions that control relevant endpoint pattern characteristics. This technique provides a facile method to generate micropatterned arrays with tailorable and controlled presentation of biological ligands for biotechnological applications and analyses of cell-material interactions.