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The Sysmex pocH-100iV Diff is an impedance hematology analyzer recently introduced for point-of-care use in veterinary practices in Europe.The purpose of this study was to validate the pocH-100iV Diff for analysis of blood samples from dogs, cats, horses, and cattle.Fresh EDTA-blood samples from healthy and ill dogs (115), cats (94), horses (91), and cattle (78) were analyzed on the pocH-100iV Diff and the Cell-Dyn 3500. Results of the automated WBC differential counts were compared with the manual differential counts for 77 dogs, 65 cats, 40 horses, and 46 cattle. HCT were compared with PCVs obtained by microhematocrit centrifugation. Furthermore, precision, linearity, carry-over, cell aging, and clinical relevance of the pocH-100iV Diff results were assessed.Most of the CBC results obtained by the pocH-100iV Diff correlated well with those of the Cell-Dyn 3500. Slightly low correlation was observed for canine MCV and hemoglobin concentration. Lymphocytes correlated well in horses and cattle, but less well in cats and dogs. The mixed cell population termed “OTHRS” (all granulocytes and monocytes for horses and cattle; neutrophils, monocytes, and basophils for cats and dogs) correlated well in all tested species. The instrument overestimated feline and canine eosinophils. In cats, platelet counts showed a strong negative bias.The overall performance of the pocH-100iV Diff was excellent with the noted limitations. The automated differential count can be used as screening tool in conjunction with evaluation of a blood smear.