Cu2+　is　closely　related　to　the　occurrence　and　development　of　Wilson＇s　disease　(WD),　and　quantitative　detection　of　various　copper　indicators　(especially　liver　Cu2　and　urinary　Cu2+)　is　the　key　step　for　the　early　diagnosis　of　WD　in　the　clinic.　However,　the　clinic　Cu2+　detection　approach　was　mainly　based　on　testing　the　liver　tissue　through　combined　invasive　liver　biopsy　and　the　ICP-MS　method,　which　is　painful　for　the　patient　and　limited　in　determining　WD　status　in　realtime.　Herein,　we　rationally　designed　a　type　of　Cu2+-activated　nanoprobe　based　on　nanogapped　gold　nanoparticles　(AuNNP)　and　poly(N-isopropylacrylamide)　(PNIPAM)　to　simultaneously　quantify　the　liver　Cu2+　content　and　urinary　Cu2+　in　WD　by　photoacoustic　(PA)　imaging　and　ratiometric　surface-enhanced　Raman　scattering　(SERS),　respectively.　In　the　nanoprobe,　one　Raman　molecule　of　2-naphthylthiol　(NAT)　was　placed　in　the　nanogap　of　AuNNP.　PNIPAM　and　the　other　Raman　molecule　mercaptobenzonitrile　(MBN)　were　coated　on　the　AuNNP　surface,　named　AuNNP-NAT@MBN/PNIPAM.　Cu2+　can　efficiently　coordinate　with　the　chelator　PNIPAM　and　lead　to　aggregation　of　the　nanoprobe,　resulting　in　the　absorption　red-shift　and　increased　PA　performance　of　the　nanoprobe　in　the　NIR-II　window.　Meanwhile,　the　SERS　signal　at　2223　cm-1　of　MBN　is　amplified,　while　the　SERS　signal　at　1378　cm-1　of　NAT　remains　stable,　generating　a　ratiometric　SERS　I2223/I1378　signal.　Both　NIR-II　PA1250　nm　and　SERS　I2223/I1378　signals　of　the　nanoprobe　show　a　linear　relationship　with　the　concentration　of　Cu2+.　The　nanoprobe　was　successfully　applied　for　in　vivo　quantitative　detection　of　liver　Cu2+　of　WD　mice　through　NIR-II　PA　imaging　and　accurate　quantification　of　urinary　Cu2+　of　WD　patients　by　ratiometric　SERS.　We　anticipate　that　the　activatable　nanoprobe　might　be　applied　for　assisting　an　early,　precise　diagnosis　of　WD　in　the　clinic　in　the　future.　©　2021　American　Chemical　Society.