To test methods used for converting observations of acoustic backscatter to estimates of the volume and transport of free gas escaping the seabed, a bubble generator has been constructed and used at sea. The bubble generator creates individual bubbles of the sizes commonly associated with methane seeps, 1-5 mm radii, which can be released at pre-planned rates. The bubble generator was deployed off the coast of New Hampshire, USA, at a depth of 55 m, and acoustic backscatter between 16-24 kHz was collected from a shipboard echo sounder while transiting over the rising bubbles. Bubble sizes and compositions (either Ar or N 2 ) were known at the source. A model for bubble evolution, accounting for changes in bubble size and composition due to hydrostatic pressure and gas diffusion across the gas-liquid boundary, was coupled with an acoustic TS model to generate predictions of the acoustic backscatter from bubbles that had risen to different depths. These predictions were then compared with experimental observation. Good agreement between prediction and observation was found in most cases, with the exception of the largest (4 mm) gas bubbles at depths of 30 m or less. The exact cause of this bias is unknown, but may be due to incorrect assumptions in models for the bubble TS, rise velocity, or mass transfer rate.