Jain N C, Paape M J, Miller R H
Department of Clinical Pathology, School of Veterinary Medicine, University of California, Davis 95616.
Am J Vet Res. 1991 Apr;52(4):630-6.
A flow cytometric method was developed to perform differential leukocyte counts on bovine blood. Blood specimens from 50 healthy Holstein cows were analyzed by use of a flow cytometer. The method entailed diluting blood with phosphate-buffered, hypotonic saline solution containing acridine orange, and performing a step-wise, 3-parameter analysis on the bases of cell size, cellular granularity, and granulocyte fluorescence. Initially, proportions of monocytes, granulocytes, and lymphocytes were determined by creating appropriate windows on dot plots of cell size (determined by forward light scatter) vs cellular granularity (determined by the logarithm of side light scatter). Eosinophils were resolved by analysis of granulocytes as dot plots of logarithms of green vs red fluorescence ascribed to acridine orange. Proportions of eosinophils and neutrophils were computed from data so generated. Microclumps of platelets spuriously affected counts of some granulocytes, particularly eosinophils. Differential leukocyte counts determined by flow cytometry generally compared favorably with those obtained by use of the conventional microscopic method, using Wright-stained blood films. Mean neutrophil and eosinophil counts determined by the 2 methods did not differ significantly, but lymphocyte counts determined by flow cytometry were significantly higher than those determined by microscopy (P less than 0.01). Correlation coefficients for counts of neutrophils, eosinophils, and lymphocytes determined by the 2 methods ranged from 0.519 to 0.833. Correlation between monocyte counts was low (r = 0.147), although mean monocyte counts determined by the 2 methods did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)
开发了一种流式细胞术方法用于对牛血液进行白细胞分类计数。使用流式细胞仪对50头健康荷斯坦奶牛的血液样本进行了分析。该方法包括用含有吖啶橙的磷酸盐缓冲低渗盐溶液稀释血液，并基于细胞大小、细胞颗粒度和粒细胞荧光进行逐步的三参数分析。最初，通过在细胞大小（由前向光散射确定）与细胞颗粒度（由侧向光散射的对数确定）的点图上创建适当的窗口来确定单核细胞、粒细胞和淋巴细胞的比例。通过将粒细胞分析为吖啶橙所致绿色与红色荧光对数的点图来分辨嗜酸性粒细胞。从所产生的数据计算嗜酸性粒细胞和中性粒细胞的比例。血小板微团会虚假地影响一些粒细胞的计数，尤其是嗜酸性粒细胞。通过流式细胞术确定的白细胞分类计数通常与使用传统显微镜方法（对经瑞氏染色的血涂片）获得的计数结果相当。两种方法确定的中性粒细胞和嗜酸性粒细胞平均计数无显著差异，但流式细胞术确定的淋巴细胞计数显著高于显微镜检查确定的计数（P小于0.01）。两种方法确定的中性粒细胞、嗜酸性粒细胞和淋巴细胞计数的相关系数范围为0.519至0.833。单核细胞计数之间的相关性较低（r = 0.147），尽管两种方法确定的单核细胞平均计数无显著差异。（摘要截短于250字）
