Technical Advance: Autofluorescence as a tool for myeloid cell analysis
Technical Advance: Autofluorescence as a tool for myeloid cell analysis
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DOI：
10.1189/jlb.0310184
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发表时间：
2010-09-01
影响因子：
5.5
通讯作者：
Chimini, Giovanna
中科院分区：
文献类型：
--
作者：
Mitchell, Andrew J.;Pradel, Lydie C.;Chimini, Giovanna
细胞AF通常被认为是流式细胞术分析的障碍。在这里，我们将AF分析复杂的白细胞混合物。使用小鼠模型，我们在多个激发和发射波长下检查了细胞AF，对具有离散模式的群体进行门控并检查表面标志物表达。在脾脏中，鉴定了所有主要的骨髓细胞群。特别是，该方法允许同时表征RPM和驻留单核细胞。当单核细胞和RPM进行了比较，RPM表现出的表型，这是参与生理过程，包括参与脂质和铁代谢的基因的表达是一致的。RPM中大量储存的三价铁的存在使得能够使用基于磁性的方法纯化这些细胞。当适用于从一系列其他器官分离的白细胞时，将AF纳入分析允许鉴定和分离生物学上重要的骨髓细胞群，包括通过常规细胞计数分析不易鉴定的亚群。J. Leukoc. 88：597-603; 2010.
Cellular AF is usually considered a hindrance to flow cytometric analysis. Here, we incorporate AF into analysis of complex mixtures of leukocytes. Using a mouse model, we examined cellular AF at multiple excitation and emission wavelengths, and populations with discrete patterns were gated and examined for surface marker expression. In the spleen, all major myeloid populations were identified. In particular, the approach allowed simultaneous characterization of RPM and resident monocytes. When monocytes and RPM were compared, RPM exhibited a phenotype that was consistent with involvement in physiological processes, including expression of genes involved in lipid and iron metabolism. The presence of large amounts of stored ferric iron within RPM enabled purification of these cells using a magnetic-based approach. When adapted for use on leukocytes isolated from a range of other organs, incorporation of AF into analysis allowed identification and isolation of biologically important myeloid populations, including subsets that were not readily identifiable by conventional cytometric analysis. J. Leukoc. Biol. 88: 597-603; 2010.