Söderhäll K, Smith V J
Dev Comp Immunol. 1983 Spring;7(2):229-39. doi: 10.1016/0145-305x(83)90004-6.
Three morphologically distinct populations of haemocytes; the granular, semigranular and hyaline cells; were isolated from the haemolymph of Carcinus maenas and other decapod crustaceans by density gradient centrifugation. Cell lysis and coagulation during separation were prevented, without significant loss of cell viability, by the use of citrate/EDTA buffer at low pH as an anticoagulant. Biochemical analyses of the haemocyte fractions revealed that prophenoloxidase is present in the granular and semigranular cells, but not the hyaline cells, and, thus, is a useful marker for cell purity in the hyaline haemocyte populations. A method for rapidly detecting prophenoloxidase contamination of the hyaline cells using L-dopa and trypsin in drop assay is described.
通过密度梯度离心法，从滨蟹及其他十足目甲壳动物的血淋巴中分离出三种形态各异的血细胞群体，即颗粒细胞、半颗粒细胞和透明细胞。在分离过程中，通过使用低pH值的柠檬酸盐/乙二胺四乙酸缓冲液作为抗凝剂，防止了细胞裂解和凝血现象，且细胞活力没有明显损失。对血细胞组分的生化分析表明，前酚氧化酶存在于颗粒细胞和半颗粒细胞中，而透明细胞中不存在，因此，它是透明血细胞群体中细胞纯度的一个有用标记物。本文描述了一种在点滴试验中使用L-多巴和胰蛋白酶快速检测透明细胞中前酚氧化酶污染的方法。
