利用相差显微镜和流式细胞术分析褶纹冠蚌血细胞类型

Analysis on the Categories of the Blood Cells from Cristaria Plicata by Phase-contrast Microscope and Flow Cytometry

利用相差显微镜对褶纹冠蚌的血细胞进行了观察,结果发现颗粒细胞和无颗粒细胞两大类。颗粒细胞还可以根据细胞和颗粒的大小分为大颗粒细胞和小颗粒细胞,无颗粒细胞也可以根据细胞的大小和细胞的核质比分为透明细胞和淋巴样细胞。另外,利用流式细胞仪进一步分析了褶纹冠蚌血细胞的类群。通过测试前向角散射光(FSC)和侧向角散射光(SSC)两个参数,分别代表细胞的大小和细胞的颗粒度,绘制双参数(SSC×FSC)点图和等高图,FSC和SSC单参数直方图,得出褶纹冠蚌血细胞可以分为两大群,即颗粒细胞(A)和无颗粒细胞(B)两种类型,其中颗粒细胞群可以划为两个亚群,即大颗粒细胞(A1)和小颗粒细胞(A2),无颗粒细胞群也可以划为两个亚群,即透明细胞(B2)和淋巴样细胞(B1)。在四种血细胞中,小颗粒细胞和透明细胞是主要的细胞类群,分别占血细胞总数的56.54%和35.90%,而大颗粒细胞和淋巴样细胞数量较少,分别为4.57%和2.99%。

Living cells were observed by phase-contrast microscope,it was considered with two types of cells：granulocytes and agranulocytes,according to the presence or the absence of granules in the cytoplasm.In addition,according to the diameter of granulocytes and granules,granulocytes were further divided into two subpopulations,large granulocytes and small granulocytes,and on the basis of the diameter of agranulocytes and N/C,agranulocytes were also further divided into two subpopulations,hyalinocytes and lymphoid hemocytes.Two parameters of light-scatter flow cytometry were set up,forward light-scatter（FSC） and side light-scatter（SSC）.FSC were given a relative indication of cell size,while SSC was an indication of complexity or granularity of cells.The analysis results by the Cellquest software were expressed as the dot and the contour plots of two parameters（FSC SSC）,the single parameter（FSC SSC） histogram of hemocytes for C.plicata.Hemocytes of C.plicata were divided into the two populations of granulocytes（A） and agranulocyts（B） by flow cytometry.The group A of granulocytes could be diveded into two sub-group,large granulocytes（A1） and small grnulocytes（A2）.The group B of agranulocytes could be also diveded into two sub-group,hyalinocytes（B2） and lymphoid hemocytes（B1）.Small granulocytes and hyalinocytes were the main cell types of C.plicata,56.54% and 35.90% in hemocyte population respectively,while large granulocytes and lymphoid hemocytes were 4.57%and2.99% respectively.