A whole-blood flow cytometric assay for leukocyte CD11b expression using fluorescence signal triggering
A whole-blood flow cytometric assay for leukocyte CD11b expression using fluorescence signal triggering
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DOI：
10.1034/j.1600-0609.2000.90198.x
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发表时间：
2000-07-01
影响因子：
3.1
通讯作者：
Hjemdahl, P
中科院分区：
文献类型：
--
作者：
Li, NL;Halldén, G;Hjemdahl, P
已开发并评价了用于测量全血中白细胞CD 11b表达的流式细胞术测定。该方法是基于荧光泛白细胞标志物RPE-CD 45触发流式细胞仪。这使得能够在全血中进行流式细胞术分析，并避免与细胞纯化和溶血相关的体外伪影。我们的方法学评价建议采用以下常规程序：用柠檬酸钠作为抗凝剂取样，在22 ℃下孵育样品，并用0.5%甲醛盐水温和固定样品。后者在24 h内提供了良好的样品稳定性。此外，该测定提供了良好的测定再现性、低标记抗体消耗和最小的样品操作（
A flow cytometric assay for measurements of leukocyte CD11b expression in whole blood has been developed and evaluated. The method is based on triggering of the flow cytometer by a fluorescent pan leukocyte marker, RPE-CD45. This enabled flow cytometric analysis in whole blood, and avoidance of in vitro artefacts related to cell purification and hemolysis. Our methodological evaluation suggested the following routine procedure: sampling with sodium citrate as the anticoagulant, sample incubation at 22 degrees C, and mild sample fixation with 0.5% formaldehyde saline. The latter provided good sample stability during 24 h. Moreover, the assay provided good assay reproducibility, low labelling antibody consumption, and minimal sample manipulation (