Deneys V, Mazzon A M, Robert A, Duvillier H, De Bruyère M
Laboratory of Immunohaematology, Catholic University of Louvain, Belgium.
Vox Sang. 1994;67(2):172-7. doi: 10.1111/j.1423-0410.1994.tb01654.x.
Reduction of leucocytes in blood components, achieved by filtration, may reduce the risk of HLA alloimmunization, virus transmission, and febrile reactions. Nevertheless, white blood cell (WBC) concentrations obtained in this way can be too low to be accurately counted with automated and manual techniques. We describe here a rapid, reliable, and very sensitive method for counting low leucocyte numbers using flow cytometry. WBC nuclei are labelled by propidium iodide. A known amount of fluorescein- and phycoerythrin-conjugated beads (Standard Brite, Coulter) is added as an indicator of the examined volume. The time required for analyzing one tube is approximately 5 min. This method was first validated by comparing WBC counts (120-2,570/microliters) assessed on a haemocytometer (Bürker) and by flow cytometry. The second step was to determine the sensitivity of the method: we diluted normal platelet concentrates with phosphate-buffered saline. The comparison of expected with observed values showed a very good correlation up to a concentration of 1 WBC/microliter. This technique is an accurate method that can be applied in blood bank quality controls and in clinical studies.
通过过滤减少血液成分中的白细胞，可能会降低HLA同种免疫、病毒传播和发热反应的风险。然而，以这种方式获得的白细胞（WBC）浓度可能过低，无法用自动化和手动技术准确计数。我们在此描述一种使用流式细胞术计数低白细胞数量的快速、可靠且非常灵敏的方法。白细胞核用碘化丙啶标记。加入已知量的荧光素和藻红蛋白偶联珠（标准亮珠，库尔特公司）作为所检测体积的指示剂。分析一管所需时间约为5分钟。该方法首先通过比较血细胞计数器（伯克氏）和流式细胞术评估的白细胞计数（120 - 2570/微升）进行验证。第二步是确定该方法的灵敏度：我们用磷酸盐缓冲盐水稀释正常血小板浓缩液。预期值与观察值的比较显示，在白细胞浓度高达1/微升时具有很好的相关性。该技术是一种准确的方法，可应用于血库质量控制和临床研究。
