Cutting-edge analysis of extracellular microparticles using ImageStream(X) imaging flow cytometry.
Cutting-edge analysis of extracellular microparticles using ImageStream(X) imaging flow cytometry.
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DOI：
10.1038/srep05237
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发表时间：
2014-06-10
影响因子：
4.6
通讯作者：
Norling LV
中科院分区：
文献类型：
--
作者：
Headland SE;Jones HR;D'Sa AS;Perretti M;Norling LV
在过去的十年中，对细胞外囊泡生物学的兴趣激增，因为这些微结构似乎被赋予了多种角色，从血液凝固到病理生理学中的细胞间通讯。为了使微粒研究发展成为临床前和临床工具，微粒水平的准确定量是一个基本要求，但它们的大小和样品流体的复杂性带来了重大的技术挑战。流式细胞术是常用的，但具有低灵敏度和准确性。使用Amnis ImageStreamX Mk II成像流式细胞仪可准确分析1 μm至20 nm范围内的校准微珠;以及微粒，可在全血、富血小板和无血小板血浆以及白细胞上清液中观察和定量。 另一个优点是所需的样品制备和体积最小。这种高通量分析仪的使用允许亲本细胞和后代微粒的表型定义以及沿着真实的时间微粒生成动力学。由于目前缺乏可靠的微粒分析技术，我们建议ImageStreamX可以有效地用于推进这一科学领域。
Interest in extracellular vesicle biology has exploded in the past decade, since these microstructures seem endowed with multiple roles, from blood coagulation to inter-cellular communication in pathophysiology. In order for microparticle research to evolve as a preclinical and clinical tool, accurate quantification of microparticle levels is a fundamental requirement, but their size and the complexity of sample fluids present major technical challenges. Flow cytometry is commonly used, but suffers from low sensitivity and accuracy. Use of Amnis ImageStreamX Mk II imaging flow cytometer afforded accurate analysis of calibration beads ranging from 1 μm to 20 nm; and microparticles, which could be observed and quantified in whole blood, platelet-rich and platelet-free plasma and in leukocyte supernatants. Another advantage was the minimal sample preparation and volume required. Use of this high throughput analyzer allowed simultaneous phenotypic definition of the parent cells and offspring microparticles along with real time microparticle generation kinetics. With the current paucity of reliable techniques for the analysis of microparticles, we propose that the ImageStreamX could be used effectively to advance this scientific field.
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DOI：
10.4049/jimmunol.1003865
发表时间：
2011-05-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Norling LV;Spite M;Yang R;Flower RJ;Perretti M;Serhan CN
通讯作者：
Serhan CN
影响因子：
6.4
作者：
Janowska-Wieczorek, A;Wysoczynski, M;Ratajczak, MZ
通讯作者：
Ratajczak, MZ
影响因子：
12.8
作者：
Foladori, Paola;Quaranta, Alberto;Ziglio, Giuliano
通讯作者：
Ziglio, Giuliano
影响因子：
4.8
作者：
Latham, Sharissa L.;Chaponnier, Christine;Combes, Valery
通讯作者：
Combes, Valery
影响因子：
11.4
作者：
Bianco, Fabio;Perrotta, Cristiana;Novellino, Luisa;Francolini, Maura;Riganti, Loredana;Menna, Elisabetta;Saglietti, Laura;Schuchman, Edward H.;Furlan, Roberto;Clementi, Emilio;Matteoli, Michela;Verderio, Claudia
通讯作者：
Verderio, Claudia