In Mono Lake, concentrations of 3 He and 4 He are greater than atmospheric equilibrium values, indicating a subsurface helium source. This assertion is supported by vertical concentration profiles that show maximum ␦ 3 He values (ϩ68%) occurring within the pycnocline. The slope of the regression between 3 He and 4 He concentrations is near 2.8 R a (R a is the atmospheric helium isotope ratio), indicating that the subsurface source has a component of mantle helium. Hydrothermal springs and gas vents from Paoha Island have similar ratios, which suggests that discharge from this thermal system is the most important source. A 3 He mass balance indicated that below the deep pycnocline at 17 m, ϳ25 cc STP of 3 He accumulates each year (ϳ90% from the influx of hydrothermal water and ϳ10% from the in situ decay of tritium). Although it is not retained because of gas transfer across the air-water interface, ϳ70 cc STP of 3 He is injected each year into the upper 17 m. When 3 He is used as a tracer, the influx of hydrothermal water below the deep pycnocline was estimated to be 0.045 Ϯ 0.025 m 3 s Ϫ1 . Because the hydrothermal water is fresher than Mono Lake, this influx causes the salinity of the monimolimnion to decrease by ϳ0.1 salinity units per year and may play a small but important role in the salinity budget of this layer, which is presently denser than and isolated from the surface water.