Preloznik-Zupan I, Cernelc P, Zontar D
Department of Haematology, University Medical Centre, Ljubljana, Slovenia.
Pflugers Arch. 2000;440(5 Suppl):R185-7.
The reticulocyte count is a clinically important indirect indicator of erythropoietic activity of the bone marrow. Reticulocyte enumeration by light microscopy is rather inaccurate and has poor reproducibility. Automation of the reticulocyte count by means of flow cytometry has considerably improved the quality of this investigation. In our study, we compared three methods of establishing the blood reticulocyte number: the microscopic brilliant cresyl blue method and two flow cytometric procedures using thiazole orange (TO), namely FACSort (Becton-Dickinson) and EPICS Profile (Coulter). The aims of the study were (1) to select the most suitable TO concentration to be used with the EPICS Profile cytometer, (2) to determine the correlation between the microscopic method and the two flow cytometric procedures, and (3) to appraise the suitability of flow cytometry for reticulocyte analysis in routine clinical work. According to our results, the most appropriate TO concentration for the EPICS Profile counter is 0.1 mg/L. We observed a good correlation between the three methods tested; the correlation coefficients ranged from 0.82 to 0.87. The mean intra-assay coefficients of variation for the microscopic method and the EPICS Profile and FACSort procedures were 27.5%, 8.4% and 6.3%, respectively.
网织红细胞计数是临床上评估骨髓红细胞生成活性的一项重要间接指标。通过光学显微镜进行网织红细胞计数相当不准确，且重复性较差。借助流式细胞术实现网织红细胞计数的自动化，极大地提高了这项检测的质量。在我们的研究中，我们比较了三种测定血液中网织红细胞数量的方法：显微镜下煌焦油蓝法以及两种使用噻唑橙（TO）的流式细胞术方法，即FACSort（贝克曼-迪金森公司）和EPICS Profile（库尔特公司）。本研究的目的是：（1）选择最适合与EPICS Profile细胞仪一起使用的TO浓度；（2）确定显微镜法与两种流式细胞术方法之间的相关性；（3）评估流式细胞术在常规临床工作中用于网织红细胞分析的适用性。根据我们的结果，EPICS Profile计数器最适宜的TO浓度为0.1 mg/L。我们观察到所测试的三种方法之间具有良好的相关性；相关系数在0.82至0.87之间。显微镜法、EPICS Profile法和FACSort法的批内变异系数均值分别为27.5%、8.4%和6.3%。
