We aimed to study lymphoma diagnostics by Cerenkov luminescence imaging (CLI). We monitored the dissemination of a spontaneous high-grade mouse lymphoma (Bc.DLFL1) in early stage; advanced stage; and terminal stage with in vivo 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) positron emission tomography (PET) / magnetic resonance imaging (MRI) and 67Ga-citrate single photon emission computed tomography (SPECT) / MRI. In vivo imaging was combined with ex vivo high resolution CLI. The use of CLI with Fluorine-18 and Gallium-67 to select infiltrated lymph nodes for tumor staging pathology was thus tested. At advanced stage, [18F]FDG PET/MRI plus ex vivo CLI allowed accurate detection of [18F]FDG accumulation in lymphoma-infiltrated tissues. At terminal stage we detected tumorous lymph nodes with SPECT/MRI and we could report the Cerenkov light emission of 67Ga. CLI with 67Ga-citrate revealed lymphoma accumulation in distant lymph node locations, unnoticeable with only MRI. Flow cytometry and immunohistochemistry confirmed these imaging results. Our study promotes the combined use of PET and CLI in preclinical studies and clinical practice. Heterogeneous [18F]FDG distribution in lymph nodes detected at sampling surgery has implications for tissue pathology processing and could direct therapy. The results with 67Ga also point to the opportunities to further apply suitable SPECT radiopharmaceuticals for CLI.