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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅴ, Jan 23-25, 2002, San Jose, USA >Immunomagnetic cell separation, imaging, and analysis using Captivate~(TM) ferrofluids
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Immunomagnetic cell separation, imaging, and analysis using Captivate~(TM) ferrofluids

机译：使用Captivate〜（TM）铁磁流体进行免疫磁性细胞分离，成像和分析

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We have developed applications of Captivate~(TM) ferrofluids, paramagnetic particles (～200 nm diameter), for isolating and analyzing cell populations in combination with fluorescence-based techniques. Using a microscope-mounted magnetic yoke and sample insertion chamber, fluorescent images of magnetically captured cells were obtained in culture media, buffer, or whole blood, while non-magnetically labeled cells sedimented to the bottom of the chamber. We combined this immunomagnetic cell separation and imaging technique with fluorescent staining, spectroscopy, and analysis to evaluate cell surface receptor-containing subpopulations, live/dead cell ratios, apoptotic/dead cell ratios, etc. The acquired images were analyzed using multi-color parameters, as produced by nucleic acid staining, esterase activity, or antibody labeling. In addition, the immunomagnetically separated cell fractions were assessed through microplate analysis using the CyQUANT Cell Proliferation Assay. These methods should provide an inexpensive alternative to some flow cytometric measurements. The binding capacities of the streptavidin-labeled Captivate ferrofluid (SA-FF) particles were determined to be 8.8 nmol biotin/mg SA-FF, using biotin-4-fluorescein, and >10~6 cells/mg SA-FF, using several cell types labeled with biotinylated probes. For goat anti-mouse IgG-labeled ferrofluids (GAM-FF), binding capacities were established to be ～0.2―7.5 nmol protein/mg GAM-FF using fluorescent conjugates of antibodies, protein G, and protein A.
机译：我们已经开发了Captivate〜（TM）铁磁流体，顺磁性颗粒（直径约200 nm）与基于荧光的技术结合用于分离和分析细胞群体的应用。使用安装在显微镜上的磁轭和样品插入室，可在培养基，缓冲液或全血中获得被磁性捕获的细胞的荧光图像，同时将非磁性标记的细胞沉淀到该室的底部。我们将这种免疫磁性细胞分离和成像技术与荧光染色，光谱学和分析相结合，以评估包含细胞表面受体的亚群，活/死细胞比率，凋亡/死细胞比率等。使用多色参数分析获得的图像由核酸染色，酯酶活性或抗体标记产生。另外，使用CyQUANT细胞增殖测定法通过微板分析来评估免疫磁性分离的细胞级分。这些方法应该为某些流式细胞仪测量提供廉价的替代方法。使用生物素-4-荧光素，链霉亲和素标记的Captivate铁磁流体（SA-FF）颗粒的结合能力确定为8.8 nmol生物素/ mg SA-FF，> 10〜6细胞/ mg SA-FF，使用几种用生物素标记的探针标记的细胞类型。对于山羊抗小鼠IgG标记的铁磁流体（GAM-FF），使用抗体，G蛋白和A蛋白的荧光偶联物将结合能力确定为〜0.2〜7.5 nmol蛋白/ mg GAM-FF。

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