Surface initiated polymerization (SIP) has become an attractive method for tailoring physical and chemical properties of surfaces for a broad range of applications. Most of those applications relied on the merit of high density coating. In this study, we explored a long overlooked field of SIP: SIP from substrates of low initiator density. We combined ellipsometry with AFM to investigate the effect of initiator density and polymerization time on the morphology of polymer coatings. In addition, we carefully adjusted the nanoscale separation of polymer chains to achieve a balance between nonfouling and immobilization capacities. We further tested the performance of those coatings on various biosensors, such as quartz crystal microbalance, surface plasmon resonance, and protein microarrays. The optimized matrices enhanced the performance of those biosensors. This report shall encourage researchers to explore new frontiers in SIP that go beyond polymer brushes.KEYWORDS: surface initiated polymerization • biosensor matrix • quartz crystal microbalance • surface plasmon resonance • protein microarray S urface initiated polymerization (SIP) was initially designed to overcome the concentration barrier problem commonly suffered in the "grafting to" strategy, so that higher grafting density (>0.06 chain nm -2 ) (1) could be achieved. As a "grafting from" strategy, SIP grew polymers from the surface tethered initiators, which could be immobilized to surfaces with high density via well-established techniques such as that of self-assembled monolayer (SAM) (2, 3). SIP has been used to tailor physical and chemical properties of surfaces for a broad range of substrates and applications. Substrates include inorganic (4-9), polymer (10-13), metal (14-17), semiconductor (18), ceramic (19), and biological materials (20)(21)(22). Applications include nonfouling (23-27), wettability (28-31), responsive surfaces (32-35), corrorison resistance (36), lithographic coating (37), colloid stability (5), and stealth effect (22, 38). Most of those applications relied on the merit of higher density coating achieved by SIP. However, we found that only SIP from a low, not high, initiator density could produce superhydrophobic surfaces (39). This finding reminded us that SIP not only produced high density brushes but also allowed us to precisely control many aspects of the resulting polymers, such as film thickness and density, composition (i.e., block copolymers), and functionality. Inspired by this finding, we decided to explore a long overlooked field of SIP: SIP from substrates of low initiator density. Specificially, we applied SIP from substrates of low initiator density to prepare biosensor matrices.The design and preparation of biosensor matrices have attracted a great deal of interest because the performance of biosensors is greatly affected by the characteristics of matrices, such as their chemical composition, three-dimensional (3D) structures, mechanical properties, and biocompatibility. For example, Liedberg et al. developed...