Donor-acceptor (D-A) conjugated polymers have large Stokes shifts, high photostability, and good biocompatibility and thus are ideal for use as Raman probes in vivo. However, few D-A conjugated polymers are used as Raman probes for Raman imaging due to their weak Raman signal intensity and intrinsic fluorescence interference. Here, we developed a D-A conjugated polymer probe (CDT-TT) containing alternating cyclopentadithiophene-thienothiophene units for Raman imaging of tumor and sentinel lymph nodes (SLNs). The CDT-TT shows a strong Raman signal at 1389 cm-1 without fluorescence and lipid background signal interference under 785 nm near-infrared light. Moreover, the CDT-TT loaded nanoparticles realized the accurate imaging of tumor cells and tumor tissues. In addition, a high-resolution margin imaging of in situ SLNs was acquired. Taken together, the established method is effective for accurate Raman detection of tumors and SLNs, which may shed new light on the development of D-A polymer Raman probes for clinical imaging.