Supported phospholipid bilayers (SPBs) are attractive substrates for biosensing applications since they support the function and integrity of many integral and membrane bound proteins, are resistant to non-specific protein adsorption, and can be functionalized with a variety of available headgroup chemistries. Through the patterning of lipid films on the micron scale, high throughput screening methodologies can be designed to study immobilized domains of biological molecules. Here, a method for creating diverse biofunctional, chemically and environmentally stable patterns based on the photoreactive phospholipids originally synthesized by David F. O'Brien is presented. Three dimensional fluidic channels are modified with polymerized phospholipid features which can be tuned to observe specific affinity binding events, such as biotin-avidin or antibody-antigen binding, while minimizing nonspecific protein binding to the area of interest. Further functionality can be introduced to the system through copolymerization of reactive phospholipids and incorporation of membrane proteins into the bilayer.