Diaphus theta is the most abundant micronektonic fish in the northwestern Pacific. Field acoustic data and biological samples were obtained both day and night off of eastern Hokkaido, Japan in September 2006 to estimate the fish density and diel changes of its vertical distribution. Differences of acoustical target strength (TS) values between at 38 and 120 kHz were applied to identify D. theta echos using field echo data. Theoretical estimation using swimbladder acoustic scattering models showed that the TS difference (TS 120-38 kHz) of larger fishes (over 60 mm) was between −4 and 1 dB; that for smaller fishes (less 60 mm) was less than .4 dB. These values differed from those of other major component species of the deep-scattering layer, such as krill and pollock, suggesting that the echo from D. theta is acoustically identifiable. Diel vertical distributions of D. theta were estimated after identification. Relatively dense schools were observed at around 400-m depth during daytime, although the schools were scattered widely above 100-m depth during nighttime. These results matched those obtained by biological sampling. In the surface layer (above 100 m), the estimated fish densities were about tenfold higher at nighttime than in daytime.