INTRODUCTION In biological researches, chicken embryos have become a classic experiment model for the exploration of the embryonic development and cell differentiation.Transferring exogenous substances into chicken embryos for producing medical antibodies has been widely used in the production practice.However, there was few studies about the diffusion effects of the different injection location on chicken embryos.The aim of this study was to explore the diffusion effects of different injection location, as well as the the impacts of different injection dosage on chicken embryo hatching rate and development, so as to provide basis and direction for the further study of chicken embryo injection.