Marti G E, Magruder L, Patrick K, Vail M, Schuette W, Keller R, Muirhead K, Horan P, Gralnick H R
Am J Hematol. 1985 Sep;20(1):41-52. doi: 10.1002/ajh.2830200107.
An interlaboratory flow cytometric comparison of several commercially available human lymphocyte subset reagents was undertaken in three different laboratories. Fresh Hypaque-Ficoll purified blood mononuclear cells were stained at 4 degrees C or 22 degrees C. Direct or indirect surface immunofluorescence was carried out at all sites using an EPICS V flow cytometer. Fullbright, 10-micron fluorescent polystyrene microspheres were used for optical alignment and standardization. A log integral fluorescent histogram gated on forward and right angle scatter was collected on 1-2 X 10(4) cells for each reagent and the proportion, of positive cell determined for each reagent. With the exception of one reagent, anti-B1, which showed an approximately twofold variation, all three laboratories showed remarkable agreement. Thus there was no significant difference noted for the following reagents: OKT4, CCT4, Leu 3a, Leu 2a, OKT8, or CCT8. We attribute these findings to the availability of quality reagents, precision instrumentation, and a standard lymphocyte preparation.
在三个不同实验室对几种市售人淋巴细胞亚群试剂进行了实验室间流式细胞术比较。用新鲜的聚蔗糖-泛影葡胺纯化的血液单核细胞在4℃或22℃下染色。所有实验点均使用EPICS V流式细胞仪进行直接或间接表面免疫荧光检测。使用10微米的全亮荧光聚苯乙烯微球进行光学校准和标准化。对每种试剂在1-2×10⁴个细胞上收集基于前向散射和直角散射的对数积分荧光直方图，并确定每种试剂阳性细胞的比例。除一种试剂抗B1显示出约两倍的差异外，所有三个实验室的结果都非常一致。因此，以下试剂之间未发现显著差异：OKT4、CCT4、Leu 3a、Leu 2a、OKT8或CCT8。我们将这些结果归因于优质试剂的可得性、精密仪器以及标准淋巴细胞制备方法。
