To provide US domestic supply of Mo-99 without using high-enriched uranium (HEU), a subcritical uranium target assembly (UTA) is irradiated by an accelerator-based neutron source to create Mo-99 through fission. Part I of this work discusses the design of a liquid lead–bismuth eutectic (LBE) windowless target for an accelerator-based neutron source development. Part II discusses how to couple this windowless target to an accelerator operating at an ultra-high vacuum and the subcritical UTA cooled by water at room temperature.Due to the windowless design of the target, the liquid LBE flow shares an ultra-high vacuum (<1.3 × 10−7 Pa or 10−9 Torr) space with the accelerator. As a result of this shared vacuum space, the LBE system must operate at a high vacuum (10−3 ∼ 10−6 Pa or 10−5 ∼ 10−8 Torr). A magnetic rotary motion feedthrough unit utilizes magnetic fluid to allow rotation of the pump while maintaining a high vacuum environment. Prior to testing the LBE system under vacuum, a pump curve measurement is performed to estimate flowrate in the system. This measurement also generates data on orifice loss coefficients, which are compared to correlations in literature. The second experiment investigates vacuum level in the LBE system during operation. High vacuum is maintained (10−3 ∼ 10−5 Pa or 10−5 ∼ 10−7 Torr) during system operation, and a residual gas analyzer (RGA) scan shows that partial pressures of residual gases in the LBE system lower over the duration of LBE system operations. The third experiment investigates the gravity driven liquid LBE flowing out of the target chamber in the return line, which is partially full. If the liquid LBE is not drained quickly enough, flooding in the target chamber could occur. The coefficient n in the Manning equation is found to be around 0.008 s/m1/3. The last experiment performed is a demonstration that a vacuum jacket could provide sufficient thermal insulation to allow coupling between 300 °C LBE loop and a water tank at room temperature. The results from these experiments have influenced the development of the neutron source for the future commercial scale Mo-99 production system.