Hemocytes Classification of Portunus trituberculatus and Oratosquilla oratoria by Imaging Flow Cytometry
[Objective]To optimize flow cytometry based methods and standards for crustacean blood cell classification.[Method]Imaging flow cytometry and microscopic observation were compared on blood cell classification of Portunus trituberculatus and Oratosquilla oratoria.[Result]Three cell types could be separated from both animals based on microscopic observation,which referred to the differences of distin-guishable intracellular particles,but the results were fluctuated by batch.Flow cytometry,based on more objective measurement of intensity and area of side scattering,could clearly divide blood cells into four populations of non-granular hemocyte(NG),small-granular hemocyte(SG),intermediate-granular hemocyte(MG),and large-granular hemocyte(LG).According to flow cytometry measurement,about 30% were non-granular hemocytes in both animals,cells with refractive structures count for the main part of whole blood cells,which was similar to that of microscopic observation results.In terms of cell size,both methods showed the same results.On one the hand,cell size increased grad-ually from non-granular hemocyte to large-granular hemocyte;On the other hand,all hemocytes of Oratosquilla oratoria were larger than those in Portunus trituberculatus.[Conclusion]Microscopic observation and flow cytometry were complementarity for each other;flow cytometry could distinguish more cell populations,was more reliable due to its objectivity,and suitable for high throughput fine analysis.
万方数据
维普
