The safety and diagnostic value of combined splenic fine-needle aspiration (FNA) and needle core biopsy (NCB) is unknown. Forty-one dogs with splenic lesions were studied prospectively. Safety was assessed in 38 dogs and no complications were encountered. Initially, clinical and anatomic pathologists reviewed each FNA and NCB sample, respectively, without knowledge of the other's results. Diagnoses were categorized as neoplastic, benign, inflammatory, normal, or nondiagnostic. The level of agreement between sampling methods was categorized as complete, partial, disagreement, or not available. Test correlation was performed in 40 dogs. Nondiagnostic results occurred in 5/40 NCB (12.5%) and no FNA samples. Neoplasia was diagnosed in 17/40 dogs (42.5%), benign changes in 20/40 dogs (50%), inflammatory disorders in 0/40 dogs, and normal 2/40 dogs (5%). One of the 40 dogs (2.5%) had a diagnosis that was equivocal for neoplasia on both tests and therefore was not categorized. Of the 35 dogs that had diagnostic samples, cytopathologic and histopathologic diagnoses agreed completely in 18/35 dogs (51.4%), partially in 3/35 dogs (8.6%), and were in disagreement in 14/35 dogs (40.0%). Pathologists collaboratively reviewed diagnoses that were in disagreement or partial agreement and altered their individual diagnoses in 6/17 dogs (35.3%) to be within partial or complete agreement, respectively. Percutaneous FNA and NCB can be performed safely in dogs with sonographic splenic changes. Results suggest that adding NCB to FNA provides complementary information in dogs with suspected splenic neoplasia. This combined protocol may improve detection of splenic neoplasia and provide neoplastic subclassification.