Li Xiao, Tibbe Arjan G J, Droog Erik, Terstappen Leon W M M, Greve Jan
Biophysical Engineering Group, University of Twente, Dienstweg 1, Building Zuidhorst, 7522 ND Enschede, The Netherlands.
Clin Vaccine Immunol. 2007 Apr;14(4):412-9. doi: 10.1128/CVI.00372-06. Epub 2007 Feb 7.
Simplification of cell enumeration technologies is necessary, especially for resource-poor countries, where reliable and affordable enumeration systems are greatly needed. In this paper, an immunomagnetic single-platform image cytometer (SP ICM) for cell enumeration based on antibody specificity is reported. A chamber/magnet assembly was designed such that the immunomagnetically labeled, acridine orange-stained cells in a blood sample moved to the surface of the chamber, where a fluorescent image was captured and analyzed for cell enumeration. The system was evaluated by applying one kind of antibody to count leukocytes and one kind for each leukocyte subpopulation: CD45 for leukocytes, CD3 for T lymphocytes, and CD19 for B lymphocytes. Excellent precision and linearity were achieved. Moreover, these cell counts, each from blood specimens of 42 to 52 randomly selected patients, were compared with those obtained by SP (TruCount) and dual-platform (DP) flow cytometry (FCM) technologies. The cell counts obtained by our system were in between those obtained from the TruCount and DP FCM methods; and good correlations were achieved (R > or = 0.95). For CD4(+) counts, as we expected, the cell count by our system was significantly higher than the CD4(+) T-lymphocyte counts obtained by SP and DP FCM methods. Immunophenotyping of the immunomagnetically selected CD4(+) cells showed that, besides CD4(+) T lymphocytes, a proportion of the CD4(+) dim monocytes was also selected. Our system is a simple immunomagnetic SP ICM, which can potentially be used for enumeration of CD3(+) CD4(+) T lymphocytes in resource-poor countries if an additional CD3 immunofluorescent label is applied.
简化细胞计数技术很有必要，尤其是对于资源匮乏的国家，这些国家急需可靠且经济实惠的计数系统。本文报道了一种基于抗体特异性的用于细胞计数的免疫磁单平台图像细胞仪（SP ICM）。设计了一种腔室/磁体组件，使得血液样本中经免疫磁标记、吖啶橙染色的细胞移动到腔室表面，在该表面捕获荧光图像并进行分析以进行细胞计数。通过应用一种抗体来计数白细胞以及针对每个白细胞亚群各用一种抗体对该系统进行了评估：用CD45计数白细胞，用CD3计数T淋巴细胞，用CD19计数B淋巴细胞。实现了出色的精密度和线性度。此外，将这些分别来自42至52名随机选择患者血液标本的细胞计数结果，与通过SP（TruCount）和双平台（DP）流式细胞术（FCM）技术获得的结果进行了比较。我们系统获得的细胞计数结果介于TruCount和DP FCM方法获得的结果之间；并且实现了良好的相关性（R≥0.95）。对于CD4(+)计数，正如我们所预期的，我们系统的细胞计数显著高于通过SP和DP FCM方法获得的CD4(+) T淋巴细胞计数。对免疫磁选的CD4(+)细胞进行免疫表型分析表明，除了CD4(+) T淋巴细胞外，还选到了一部分CD4(+)弱阳性单核细胞。我们的系统是一种简单的免疫磁SP ICM，如果应用额外的CD3免疫荧光标记，它有可能用于资源匮乏国家的CD3(+) CD4(+) T淋巴细胞计数。
