Assessment of Flow Cytometry in Counting Blood of Whale Sharks as a Rapid and Reliable Method

Konomi Ito; Sawsan Ghattas; Makio Yanagisawa; Senzo Uchida; Hiroki Sakail; Tokuma Yanail

Volume : III, Issue : XII, December - 2014

Assessment of Flow Cytometry in Counting Blood of Whale Sharks as a Rapid and Reliable Method

Konomi Ito, Sawsan Ghattas, Makio Yanagisawa, Senzo Uchida, Hiroki Sakail, Tokuma Yanail

Abstract :

Three captive whale sharks were used for blood collection. A CBC was performed manually based on Natt and Herrick�s method and a Neubauer hemacytometer. Leukocyte differential counts were measured on M-G stained blood smear. Cytochemical stains for reukocytes were conducted with MPR, PAS and SBB. The CBC were established, total reukocyte count. Granulocytes were identified heterophilic, eosinophilic and basophilic granulocytes. Heterophilic granulocyte and lymphocyte were predominant leukocytes. Eosinophilic granulocytes were positive with all three cytochemical stains. Heterophilic granulocytes showed weak reactivity with MPR, and eccentrically-located reactivity with PAS reaction. Basophilic granulocytes were only positive with PAS. With DiOC stain, several blood cell populations were identified by different FL-1, FSC and SSC properties. FL-1 vs. SSC or FSC vs. SSC dot-plot of stained blood cells showed four separate cell populations, namely erythrocytes, a mixture of three types of granulocytes, lymphocytes plus thrombocytes and monocytes plus immature cells. Each leukocyte percentages counted by the FACS analysis had relatively good correlation with those counted microscopically. Finally, in whale sharks, blood examination based on flow cytometric analysis may be useful for clinical diagnose as quick and reliable hematological technique.