Abbott D L, McGrath J P
Toxicology Division, Division of Eli Lilly & Co, Greenfield, IN 46140.
Am J Vet Res. 1991 May;52(5):723-7.
An automated reticulocyte counting method that used a flow cytometer and the nucleic acid staining dye, thiazole orange, was developed. Anticoagulated (EDTA) blood specimens were suitable for flow cytometric reticulocyte counting when stored at 4 C for 96 hours after collection. Thiazole orange-stained samples were stable for 5.5 hours after staining when stored capped at 20 C and protected from light. Flow cytometric and manual microscopic reticulocyte counts were compared for counts in the 0.27 to 5.32% range (as determined by flow cytometry) and 0.10 to 4.90% range (as determined by 1 technician). Although the results of flow cytometric analysis generally correlated well (r = 0.821) with manual counts, there was poor correlation between the procedures for counts less than or equal to 2.0% (r less than or equal to 0.272). Linearity of flow cytometric counts over the range 0.27 to 14.46% was excellent (r = 0.999). Within-run precision of flow cytometric counts (% coefficient of variation [cv] = 3 to 5) was superior to manual microscopic counts obtained by one technician (% cv = 19 to 23) and to manual microscopic counts, which were an average of counts done by 3 technicians (% cv = 8 to 18). Comparable flow cytometric counts were obtained by counting 50,000 or 100,000 blood cells in the flow cytometer.
开发了一种使用流式细胞仪和核酸染色染料噻唑橙的自动网织红细胞计数方法。抗凝（乙二胺四乙酸）血液标本在采集后于4℃储存96小时后适用于流式细胞网织红细胞计数。噻唑橙染色的样本在20℃加盖储存并避光时，染色后5.5小时内稳定。比较了流式细胞术和手工显微镜网织红细胞计数，计数范围为0.27%至5.32%（由流式细胞术测定）和0.10%至4.90%（由一名技术人员测定）。尽管流式细胞分析结果与手工计数总体相关性良好（r = 0.821），但计数小于或等于2.0%时，两种方法之间的相关性较差（r小于或等于0.272）。流式细胞计数在0.27%至14.46%范围内的线性极佳（r = 0.999）。流式细胞计数的批内精密度（变异系数百分比[cv]=3至5）优于一名技术人员进行的手工显微镜计数（cv%=19至23）以及3名技术人员平均计数的手工显微镜计数（cv%=8至18）。通过在流式细胞仪中计数50,０００或100,０００个血细胞可获得可比的流式细胞计数结果。
