The feasibility of 2DCOS (two-dimensional correlation spectroscopy) in the quality inspection of surimi was tested in this study.The changes of silver carp surimi protein secondary structure induced by freeze-thaw treatment were studied in suit using 2DCOS combined with ATR-MIR (attenuated total reflectance mid-infrared). The results showed that freeze-thaw cycles accelerated the oxidation process of surimi protein. The content of α-helix and intramolecular aggregated β-sheet all decreased, however the content of the anti-parallel β-sheet increased. 2DCOS analysis under the external perturbation of freeze-thaw cycles showed the change order was: α-helix, intramolecular aggregated β-sheet→antiparallel β-sheet→carbonyl. 2DCOS analysis under the perturbation of temperature showed that the protein secondary structure has been seriously damaged after three freeze-thaw cycles. And the effect of temperature on the carbonyl group of proteins was much smaller than that of the freeze-thaw cycles. These results indicated that the 2DCOS could explore and reflect the change degree of surimi protein. If these changes can be quantified, the 2DCOS could be used to detect the freshness of surimi.