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A real-time wireless sensor network platform capable of maintaining lossless data transmission over several minutes of continuous, high-rate sampling is presented in this paper. The platform was designed specifically to provide the capability to enable expeditious system identification, as well as load rating of highway bridges without compromising the typical data acquisition parameters employed in comparable cable-based tests. Consequently, the hardware signal conditioning interface permits data collection from a variety of sensors typical to structural health monitoring, including accelerometers, strain transducers, and temperature sensors. The embedded software features a proprietary network transmission protocol capable of lossless, real-time delivery of up to 40 measurement channels at an effective sampling rate of 128 samples per second per channel. Documented in this paper is a field study on an end-of-service highway bridge in which ambient vibration monitoring was performed using 60 accelerometers interfaced with 30 wireless sensor nodes operating within one of two simultaneously operating star topology networks. In addition, an experimental load rating of the entire structure was performed through large-scale strain measurement facilitated by the same wireless sensor network platform.