Objective To separate and purify intrahepatic macrophages from Microtus fortis Mf and identify its phagocy?tosis. Methods The intrahepatic macrophages from Mf were separated and purified by perfusion collagenase digestion and density gradient centrifugation. The function of the cells was identified by FACS analysis and ink phagocytosis activity. Results The macrophage cells from the liver of Mf were obtained. These cells were bright and circular and grew adhering to the wall. The proportion of the living cells was 95%. The binding rate of these cells from Mf with anti?mouse CD14 antibody Clone Sa2?8 was about 50%of the rate of macrophage from C57BL/6 mice with this monoclonal antibody. The result of ink?phagocytosis ex?periment of macrophage cells from the liver of Mf was positive. Conclusion The method above mentioned is useful to separate and purify macrophage from the liver of Mf. The study builds the foundation for further research on macrophages of Mf against Schistosoma japonicum.