Neonatologists use immature granulocytes (IG) in manual differential counts as an indicator of sepsis. This study was designed to compare the predictive ability of automated vs manual IG counts for neonatal sepsis. Infants undergoing sepsis evaluation were identified prospectively for study if a CBC count was obtained in temporal proximity to the blood culture. Automated IG counts were obtained from the research software of the Sysmex XE-2100 (Sysmex, Kobe, Japan). Manual average IG counts were obtained from two 100-cell manual differential counts independently performed by a technologist and a hematopathology resident. A comparative analysis of manual and automated IG counts showed considerable overlap of ranges. The highest positive blood culture rate occurred in the nonneutropenic preterm subset of infant older than 7 days (21/55 [38%]). For these infants, elevated IG counts by manual and automated methods were associated significantly with positive blood culture results (odds ratio, manual, 3.74; odds ratio, automated, 3.63), albeit with low sensitivity.