Reticulocyte count using thiazole orange. A flow cytometry method.
Reticulocyte count using thiazole orange. A flow cytometry method.
复制标题
使用噻唑橙进行网织红细胞计数。
DOI：
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
R. Verwilghen
中科院分区：
文献类型：
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作者：
L. V. D. hove;Willy Goossens;Van Duppen;R. Verwilghen
最近，流式细胞术技术已被开发出来，以取代显微镜网织红细胞计数。我们使用噻唑橙子，一种RNA结合荧光染料，区分网织红细胞和成熟红细胞。评价了噻唑橙子和Retic-Risk软件包在不同流量仪器上的常规分析性能。所应用的方法学以容易进行的方式半自动地分析了10（4）个细胞。用EDTA二钾抗凝血（静脉注射后稳定30 h）获得了一致的结果，噻唑橙子溶液中的孵育时间在25 ℃下为2 - 7 h。这使得样本采集和储存以及测定性能具有灵活性，结果无变化。孵育温度的变化高达30摄氏度没有可测量的影响。对于正常、低和高网织红细胞计数，所得值显示出良好的线性、精密度和准确度。然而，观察到干扰：RBC自发荧光、有核RBC、Howell-Jolly小体、高白细胞计数、高血小板计数和巨血小板，所有这些都假性增加了网织红细胞数量。通过软件门校正消除了这些伪影，从而留下不到5%的样品通过显微镜方法重新分析。噻唑橙子流式细胞术方法被确定为一种快速，可靠的方法，用于常规临床定量网织红细胞。
Recently flow cytometry techniques have been developed to replace the microscope reticulocyte count. We used thiazole orange, a RNA binding fluorochrome, to discriminate reticulocytes from mature erythrocytes. Thiazole orange and the Retic-COUNT software package were evaluated for performance of routine analysis on different flow instruments. The applied methodology analysed 10(4) cells semi-automatically in an easily performed manner. Consistent results were obtained with dipotassium EDTA anticoagulated blood (stable for 30 h after venesection), with incubation times in thiazole orange solution ranging from 2 to 7 h at 25 degrees C. This allowed flexibility in specimen collection and storage and assay performance with no change in results. Changes of incubation temperature up to 30 degrees C had no measurable effect. The values obtained showed good linearity, precision and accuracy for normal, low and high reticulocyte counts. However interferences were observed: RBC autofluorescence, nucleated RBC, Howell-Jolly bodies, high leucocyte count, high platelet count and giant platelets, all falsely increased the number of reticulocytes. These artifacts were eliminated by software gate corrections, thus leaving less than 5% of the specimen to be reanalysed by the microscopic method. The thiazole orange flow cytometric method was determined to be a fast, reliable method for the routine clinical quantitation of reticulocytes.
样本量对网织红细胞计数和粪便检查的影响。
影响因子：
4.6
作者：
Greenberg,ER;Beck,JR
通讯作者：
Beck,JR