In this paper, we constructed a label-free ratiometric fluorescence nanoprobe for ascorbic acid (AA) on the basis of the change in dual-emission signals by the specific reaction between cobalt oxyhydroxide (CoOOH) and AA. CoOOH nanoflakes were used as the recognition unit while polymer dots (PFO dots) and gold nanoclusters (AuNCs) served as luminescent units of the nanoprobe. After electrostatic assembly of PFO dots and AuNCs on the surface of CoOOH nanoflakes, the energy transfer between the PFO dots and CoOOH could occur and thereby quench the fluorescence of the PFO dots. When AA was present, CoOOH nanoflakes were reduced to Co²⁺, resulting in a recovery of the fluorescence of the PFO dots. In the meantime, the generated Co²⁺ could combine with the surface ligands of AuNCs, accompanied by a reduction in the luminescence intensity of the AuNCs. Therefore, the fluorescence intensity ratio of PFO dots to AuNCs would be related to the concentration of AA. The nanoprobe enabled highly sensitive analysis of AA with a detection limit of 1.9 μM and showed great performance in human serum samples and living cells. This report may not only endow a CoOOH-based nanoprobe with improved ability but also offer a novel strategy for the construction of ratiometric nanoprobes for AA and have potential applications in research into and clinical diagnosis of AA-related diseases.