【摘要】 [目的]比较优化适用于多种甲壳动物血细胞分析的流式方法和分类标准。[方法]对比分析了图像流式细胞术和显微观察法在三疣梭子蟹和口虾蛄2种海产甲壳动物血细胞分类中的应用。[结果]显微观察参考可辨的胞内颗粒，从2种动物血细胞中区分出无颗粒细胞、小颗粒细胞及大颗粒细胞3种，但分类标准主观性较强，批次统计波动相对较大；流式测量以更加客观的侧向散射强度和面积测量值为标准，将血细胞清晰分为无颗粒、小颗粒、中颗粒和大颗粒细胞4类，其中无颗粒细胞占比均在30%左右，含有颗粒的细胞占比较大，总体与显微观察结果相近。从细胞大小来看，2种方法呈现出一致的分析结果，一方面从无颗粒细胞到大颗粒细胞，呈现逐渐增大的特征；另一方面，口虾蛄的各类血细胞均大于三疣梭子蟹的血细胞。[结论]2种方法评价标准既有区别又有联系，因此结果相近；在适用范围方面具有一定的互补性，流式测量能区分4类细胞，而且通量高，适合大量样本的精细分析。更多还原

【Abstract】 [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 distinguishable 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 gradually 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.更多还原