The Juvenile Salmon Acoustic Telemetry System (JSATS) is an active sensing technology developed by the U.S. Army Corps of Engineers, Portland District, for detecting and tracking small fish. It is used primarily for evaluating behavior and survival of juvenile salmonids migrating through the Federal Columbia River Power System to the Pacific Ocean. It provides critical data for salmon protection and development of more “fish-friendly” hydroelectric facilities. The objective of this study was to design and build a Measurement and Calibration System (MCS) for evaluating the JSATS components, because the JSATS requires comprehensive acceptance and performance testing in a controlled environment before it is deployed in the field. The MCS consists of a reference transducer, a water test tank lined with anechoic material, a motion control unit, a reference receiver, a signal conditioner and amplifier unit, a data acquisition board, MATLAB control and analysis interface, and a computer. The fully integrated MCS has been evaluated successfully at various simulated distances and using different encoded signals at frequencies within the bandwidth of the JSATS transmitter. The MCS provides accurate acoustic mapping capability in a controlled environment and automates the process that allows real-time measurements and evaluation of the piezoelectric transducers, sensors, or the acoustic fields. The MCS has been in use since 2009 for acceptance and performance testing of, and further improvements to, the JSATS.