Three types of granulocytes were observed in peripheral blood of Japanese angelshark Squatina japonica (Squatinidae, Squatiniformes, Squalimorphii, Elasmobranchii). The neutrophil was the only phagocytic granulocyte and had two types of granules (neutrophil granule, NG； type A, NG-A； type B, NG-B) with stratified structure. The NG-A was long-elliptic shape, and consisted of three layers: Eosinophilic rod-shaped inner layer (L0), eosinophilic middle layer (L1) with long-spindle outline, and basophilic outer layer (L2). The NG-B was round or oval, and both inner (L0) and outer (L1) layers of this granule was chromophobic. Several lysozomal enzymes were detected in NG-B, but the positive site was different among enzymes. Alpha-naphtyl acetate esterase, α-naphtyl butyrate esterase and naphthol AS-D chloroacetate esterase (NASDCAE) were detected in the L0 of NG-B. According to the acid phosphatase positive site, NG-B were classified into three types: NG-B with positive L0 (negative L1), NG-B with negative L0 (positive L1) and NG-B with positive L0 (strongly) and L1. Furthermore, the L0 of NG-B around nucleus showed positive reaction to periodic acid Schiff reaction after digestion with α-amylase. The neutrophils lacked alkaline phosphatase, β-glucuronidase and peroxidase. The L0 and L1 of NG-A showed positive reaction to Sudan black B and NASDCAE.