The use of spinning-disk confocal microscopy for the intravital analysis of platelet dynamics in response to systemic and local inflammation.
The use of spinning-disk confocal microscopy for the intravital analysis of platelet dynamics in response to systemic and local inflammation.
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DOI：
10.1371/journal.pone.0025109
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Kubes P
中科院分区：
文献类型：
--
作者：
Jenne CN;Wong CH;Petri B;Kubes P
血小板是炎症的核心参与者，是先天免疫反应的重要组成部分。在活体宿主中可视化血小板的能力对于理解它们在这些过程中的作用至关重要。过去的方法包括过继转移标记的血小板、非特异性染料或使用荧光抗体在体内标记血小板。通常，这些技术导致血小板的活化或特异性血小板受体的阻断。在这份报告中，我们描述了两种新的方法，血小板生物学的活体可视化，静脉注射标记的抗CD 49 b，标记所有的血小板，和CD 41-YFP转基因小鼠，其中一个百分比的血小板表达YFP。这两种方法标记内源性血小板，并允许其可视化使用旋转盘共聚焦荧光显微镜。在LPS诱导的炎症之后，我们能够测量在许多不同组织的脉管系统内观察到的血小板聚集体的数量和大小的显著增加。对这些血小板聚集体的实时观察显示，它们是不断膨胀并脱落进入循环的大型动态结构。使用这些技术，我们描述了第一次，血小板募集，并在小鼠的许多组织内的行为，无论是在控制条件下和LPS诱导的炎症。
Platelets are central players in inflammation and are an important component of the innate immune response. The ability to visualize platelets within the live host is essential to understanding their role in these processes. Past approaches have involved adoptive transfer of labelled platelets, non-specific dyes, or the use of fluorescent antibodies to tag platelets in vivo. Often, these techniques result in either the activation of the platelet, or blockade of specific platelet receptors. In this report, we describe two new methods for intravital visualization of platelet biology, intravenous administration of labelled anti-CD49b, which labels all platelets, and CD41-YFP transgenic mice, in which a percentage of platelets express YFP. Both approaches label endogenous platelets and allow for their visualization using spinning-disk confocal fluorescent microscopy. Following LPS-induced inflammation, we were able to measure a significant increase in both the number and size of platelet aggregates observed within the vasculature of a number of different tissues. Real-time observation of these platelet aggregates reveals them to be large, dynamic structures that are continually expanding and sloughing-off into circulation. Using these techniques, we describe for the first time, platelet recruitment to, and behaviour within numerous tissues of the mouse, both under control conditions and following LPS induced inflammation.
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DOI：
10.1097/00005373-200203000-00005
发表时间：
2002-03-01
影响因子：
--
作者：
Fujimi, S;Ogura, H;Sugimoto, H
通讯作者：
Sugimoto, H
影响因子：
20.3
作者：
Andonegui, G;Kerfoot, SM;Kubes, P
通讯作者：
Kubes, P
影响因子：
9.5
作者：
Kaelsch, Thorsten;Elmas, Elif;Dempfle, Carl-Erik
通讯作者：
Dempfle, Carl-Erik
影响因子：
30.5
作者：
Lee, Woo-Yong;Moriarty, Tara J.;Kubes, Paul
通讯作者：
Kubes, Paul
影响因子：
82.9
作者：
Falati, S;Gross, P;Furie, B
通讯作者：
Furie, B