Separation of the Haemocyte Populations of Carcinus Maenas and Other Marine Decapods, and Prophenoloxidase Distribution

Three morphologically distinct populations of haemocytes; the granular, semigranular and hyaline cells; were isolated from the haemolymph of Carcinus maenas and other decapod crustaceans by density gradient centrifugation. Cell lysis and coagulation during separation were prevented, without significant loss of cell viability, by the use of citrate/EDTA buffer at low pH as an anticoagulant. Biochemical analyses of the haemocyte fractions revealed that prophenoloxidase is present in the granular and semigranular cells, but not the hyaline cells, and, thus, is a useful marker for cell purity in the hyaline haemocyte populations. A method for rapidly detecting prophenoloxidase contamination of the hyaline cells using L-dopa and trypsin in drop assay is described.