In order to study the distribution pattern of chloride cells (CCs) on the gills of the juvenile Acipenser sinensis, immune localization of CCs was studied by using the methods of HE and immunofluorescence staining in this paper. The results indicated that the optimal dilution of the first antibody α5 and the fluorescent second antibody Alexa 488 donkey-anti-mouse was 1:1 500 and 1:500, respectively, which had good effect on the location of Na⁺-K⁺-ATPase immunoreactive cells (NKA-IR cells, i.e., CCs) and could be used to locate the CCs on the gills of juvenile A. sinensis. The number of the CCs on gill filaments (4.72 ± 0.34 ind/100 μm) was significantly (P<0.001) greater than that on gill lamella (0.64±0.19 ind/100μm).The mean area of the CCs had no significant difference (P>0.05) between gill filaments (187.28 ± 18.30 μm²) and lamella (159.53 ± 16.72 μm²). The shape factor of the CCs in the gill filaments and gill lamella was 0.72 ± 0.33 and 0.68 ± 0.46, respectively, which showed no significant difference (P>0.05) between them. Thus, the CCs of juvenile A. sinensis were oval and mostly found on the gill filament, especially at the base, with a small amount on gill lamella. The CCs on the gill filament and lamella were of the same size.