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Immunocompetence is an important parameter that reflects disease resistance in fish. Very few methods to examine immunocompetence in vivo have been developed, even for mammals. In the present study, we present a unique method for analyzing local immune responses using fish fin. We first demonstrated the migration of granulocytes to the site of zymosan injection in fin in a dose-dependent manner and that this could be easily observed macroscopically due to the fin membrane transparency. We also demonstrated phagocytic activity of accumulated leukocytes after zymosan administration and that almost all phagocytes were granulocytes. In addition, we succeeded to detect respiratory burst activity of granulocytes in vivo without any in vitro treatment of cells, indicating that our present method is suitable for the analysis of granulocyte phagocytic function in vivo. The method provides a unique tool applicable for fishes that possess transparent fins and may lead to better understanding of the mechanisms of local immune responses in fish.A simple and unique method for analyzing local immune responses using fish fin in vivo was developed.Migration of granulocytes to the site of zymosan injection in fin was observed in a dose-dependent manner.Migration of granulocytes was easily observed macroscopically due to transparency of the fin membrane.In vivo phagocytic activity of granulocytes that migrated to the fin was observed after zymosan administration.Respiratory burst activity of granulocytes at the fin was also detected in vivo.