Monoclonal antibodies specific for hemocyte sub-populations of the European flat oyster, Ostrea edulis, were prepared using separated granulocytes and hyalinocytes as antigen sources. Six monoclonal antibodies specific for hemocytes were selected. Five of them showed a specificity for more than one hemocyte type and one hybridoma produced a monoclonal antibody reacting specifically with granulocytes. At the ultrastructural level, this monoclonal antibody demonstrated epitopes principally in dense cytoplasmic granules of granulocytes. Western blotting analysis indicated that a peptide of 50 kDa was recognized by this antibody. It was therefore used to investigate granulocyte distribution and ontogenesis in European flat oysters using immunohistochemistry. Granulocytes were mostly observed in connective tissues in different organs. Their distribution pattern in digestive gland, mantle, gills and gonad may indicate different functional status. Morover, the absence of granulocytes in early larval stages can partly correspond to an immature immune system in oyster larvae. (C) 2001 Elsevier Science Ltd. All rights reserved.