A calcium (Ca) prerinse before a fluoride (F) rinse has been shown to increase oral F levels. We tested the anticaries effect of this combination in a dose-response in situ caries model. In a double-blind, crossover experiment, 10 volunteers carried enamel slabs in palatal appliances for 14 days, during which they rinsed twice/day with one of four rinse combinations: (1) a placebo prerinse (150 mM sodium lactate) followed by a distilled water rinse (negative control); (2) a placebo prerinse followed by a 250 ppm F rinse; (3) a placebo prerinse followed by a 1,000 ppm F rinse, or (4) a Ca prerinse (150 mM Ca, as calcium lactate) followed by a 250 ppm F rinse. Sucrose solution was dripped onto the slabs 8×/day to simulate a high cariogenic challenge. The percent surface hardness loss (%SHL) was significantly lower in the Ca prerinse used with the 250 ppm F rinse group (%SHL = 38.0 ± 21.0) when compared with the F rinse alone (%SHL = 59.5 ± 24.1) and similar to the 1,000 ppm F rinse group (%SHL = 42.0 ± 18.3). Compared with the 250 ppm F rinse, the Ca prerinse increased biofilm fluid F only twice (nonsignificant). However, it greatly increased F in biofilm solids (∼22×). The Ca prerinse had little effect on loosely or firmly bound enamel F. The results showed an increased level of protection against demineralization by the use of a Ca prerinse, which seems to be caused by the enhancement of F concentration in the biofilm.