A new method for fish leucocyte counting and partial differentiation by flow cytometry

Author: Inoue T.   Moritomo T.   Tamura Y.   Mamiya S.   Fujino H.   Nakanishi T.  

Publisher: Academic Press

ISSN: 1050-4648

Source: Fish & Shellfish Immunology, Vol.13, Iss.5, 2002-11, pp. : 379-390

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Abstract

A simple and rapid method for analysis of fish blood cells is presented. Carp (Cyprinus carpio) blood was diluted 200 times with Hanks' solution containing 1 µg/ml of DiOC6(3) which is a fluorescent, lipophilic dye. After staining for 10 min, the blood cells were measured by a flow cytometer (FACS). Several blood cell populations were identified by different FL-1 (green fluorescence), FSC (forward scatter), and SSC (side scatter) properties. FL-1 v. SSC or FSC v. SSC dot-plot of stained blood cells displayed five separate cell populations: erythrocytes; a mixture of thrombocytes plus lymphocytes; monocytes; neutrophils; and basophils. The number of each type of blood cell counted by the FACS was in good agreement with those counted microscopically. .