Tibbe Arjan G J, de Grooth Bart G, Greve Jan, Rao Chandra, Dolan Gerald J, Terstappen Leon W M M
Biophysical Techniques Group, Faculty of Applied Physics, Twente University, PO Box 217, 7500 AE Enschede, The Netherlands.
Cytometry. 2002 Mar 1;47(3):173-82. doi: 10.1002/cyto.10061.
A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent-labeled cells and beads. Materials and Methods System performance was demonstrated with 6-microm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4-APC, CD8-Cy5.5, and CD14-APC/Cy7 in whole blood.
The sensitivity of the system was demonstrated using APC-labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm(2) and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three-color CD4-CD8-CD14 assay.
We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD-technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation.
开发了一种细胞分析系统，用于对从全血中选出的磁性排列细胞进行计数和区分。提取的细胞信息类似于从光盘（CD）读取的音乐信息。在此，我们描述该系统的光学设计和数据处理。使用荧光标记的细胞和珠子展示了该系统的性能。材料与方法 使用标记有磁性纳米颗粒和别藻蓝蛋白（APC）的6微米聚苯乙烯珠子以及全血中经免疫磁性排列、用恶嗪750和CD4 - APC、CD8 - Cy5.5以及CD14 - APC/Cy7进行荧光标记的白细胞来展示系统性能。
使用APC标记的珠子展示了该系统的灵敏度。借助此系统，含有333个APC分子的珠子能够轻松从背景中分辨出来。这种灵敏度水平是商用流式细胞仪无法达到的。在0.6米的镍线之间，最多可对20,000个经免疫磁性标记的细胞进行排列和分析，这些镍线分布在18平方毫米的表面积上，且从取决于腔室高度的血容量中提取细胞。通过进行三色CD4 - CD8 - CD14检测展示了该系统的实用性。
我们构建了一种基于免疫磁性细胞选择、细胞排列以及采用CD技术分析荧光信号的细胞分析系统，其性能与当前商用分析仪相当或更优。细胞分析可在全血或任何其他类型的细胞悬液中进行，无需进行大量样本制备。
