Imaging Flow Cytometry Elucidates Limitations of Microparticle Analysis by Conventional Flow Cytometry
Imaging Flow Cytometry Elucidates Limitations of Microparticle Analysis by Conventional Flow Cytometry
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DOI：
10.1002/cyto.a.22494
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发表时间：
2014-09-01
期刊：
影响因子：
3.7
通讯作者：
Lannigan, Joanne
中科院分区：
文献类型：
--
作者：
Erdbruegger, Uta;Rudy, Christine K.;Lannigan, Joanne
微粒 (MP) 是响应激活、细胞损伤或凋亡而从细胞膜释放的亚微米囊泡。尽管尚无标准化的测量方法，但 MP 的临床重要性已得到越来越多的认识。流式细胞术 (FCM) 是最常用的技术，然而，由于 MP 尺寸较小，以及当前 FCM 仪器的限制，该方法的准确鉴定受到影响。我们决定研究将 FCM 与成像相结合（例如 ImagestreamX 成像 FC (ISX)）的使用是否是表征 MP 的更灵敏的方法。将 FCM 与成像相结合消除了传统 FCM 所表现出的一些局限性，同时还提供形态学确认以及区分真正的单个事件与聚集体和细胞碎片的能力。与 ISX 相比，标准非专用 FCM 的检测限不是最佳的。评估低于 0.200 毫米的 MP 和确定尺寸仍然是一个挑战，因为一些 MP 仍低于 ISX 的检测限。标准化校准器需要进一步开发，以更准确地反映议员的身体特征。 (C) 2014 年国际细胞计数促进会
Microparticles (MPs) are submicron vesicles released from cell membranes in response to activation, cell injury, or apoptosis. The clinical importance of MPs has become increasingly recognized, although no standardized method exists for their measurement. Flow cytometry (FCM) is the most commonly used technique, however, because of the small size of MPs, and the limitations of current FCM instrumentation, accurate identification is compromised by this methodology. We decided to investigate whether the use of FCM combined with imaging, such as is possible with the ImagestreamX imaging FC (ISX), would be a more sensitive approach to characterizing MPs. Combining FCM with imaging eliminates some of the limitations demonstrated by conventional FCM, whereas also providing morphological confirmation and the ability to distinguish true single events from aggregates and cell debris. The detection limit of standard nonspecialized FCM is suboptimal when compared to ISX. Evaluating MPs below 0.200 mm and sizing remain a challenge as some MPs remain below the detection limit of ISX. Standardized calibrators, that more closely reflect the physical characteristics of MPs, need further development. (C) 2014 International Society for Advancement of Cytometry