Although the ability to measure vertical eddy fluxes of gases from aircraft platforms represents an important capability to obtain spatially resolved data, accurate and reliable determination of the turbulent vertical velocity presents a great challenge. A nine-hole hemispherical probe known as the “Best Air Turbulence Probe” (often abbreviated as the “BAT Probe”) is frequently used in aircraft-based flux studies to sense the airflow angles and velocity relative to the aircraft. Instruments such as inertial navigation and global positioning systems allow the measured airflow to be converted into the three-dimensional wind velocity relative to the earth’s surface by taking into account the aircraft’s velocity and orientation. Calibration of the aircraft system has previously been performed primarily through in-flight experiments, where calibration coefficients were determined by performing various flight maneuvers. However, a rigorous test of the BAT Probe in a wind tunnel has not been previously undertaken. The authors summarize the results of a complement of low-speed wind tunnel tests and in-flight calibrations for the aircraft–BAT Probe combination. Two key factors are addressed in this paper: The first is the correction of systematic error arising from airflow measurements with a noncalibrated BAT Probe. The second is the instrumental precision in measuring the vertical component of wind from the integrated aircraft-based wind measurement system. The wind tunnel calibration allows one to ascertain the extent to which the BAT Probe airflow measurements depart from a commonly used theoretical potential flow model and to correct for systematic errors that would be present if only the potential flow model were used. The precision in the determined vertical winds was estimated by propagating the precision of the BAT Probe data (determined from the wind tunnel study) and the inertial measurement precision (determined from in-flight tests). The precision of the vertical wind measurement for spatial scales larger than approximately 2 m is independent of aircraft flight speed over the range of airspeeds studied, and the 1σ precision is approximately 0.03 m s-1.