Laser Doppler velocimetry (LDV) based on a differential laser Doppler system has been widely used in fluid mechanics to measure particle velocity. However, the two outgoing lights must intersect strictly at the measurement position. In cross-interface applications, due to interface effects, two beams of light become easily disjointed. To address the issue, we present a laser velocimeter in a coaxial arrangement consisting of the following components: a single-frequency laser (wavelength λ = 532 nm) and a Twyman–Green interferometer. In contrast to previous LDV systems, a laser velocimeter based on the Twyman–Green interferometer has the advantage of realizing cross-interface measurement. At the same time, the sensitive direction of the instrument can be changed according to the direction of the measured speed. We have developed a 4000 m level laser hydrothermal flow velocity measurement prototype suitable for deep-sea in situ measurement. The system underwent a withstand voltage test at the Qingdao Deep Sea Base, and the signal obtained was normal under a high pressure of 40 MPa. The velocity contrast measurement was carried out at the China Institute of Water Resources and Hydropower Research. The maximum relative error of the measurement was 8.82% when compared with the acoustic Doppler velocimeter at the low-speed range of 0.1–1 m/s. The maximum relative error of the measurement was 1.98% when compared with the nozzle standard velocity system at the high-speed range of 1–7 m/s. Finally, the prototype system was successfully evaluated in the shallow sea in Lingshui, Hainan, with it demonstrating great potential for the in situ measurement of fluid velocity at marine hydrothermal vents.