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穿刺伤口止血和蒙太奇广域电子显微镜分析样品的制备
穿刺伤口止血和蒙太奇广域电子显微镜分析样品的制备
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JoVE Journal Medicine
Puncture Wound Hemostasis and Preparation of Samples for Montaged Wide-Area Electron Microscopy Analysis

穿刺伤口止血和蒙太奇广域电子显微镜分析样品的制备

Full Text
621 Views
05:41 min
May 24, 2024

DOI: 10.3791/66479-v

Kelly Ball1, Irina Pokrovskaya1, Brian Storrie1

1Department of Physiology and Cell Biology,University of Arkansas for Medical Sciences

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Please note that some of the translations on this page are AI generated. Click here for the English version.

本文将介绍止血血栓形成的穿刺伤口操作方法。形成的血栓很大,直径为数百微米。因此,体积成像方法是合适的。我们建议将蒙太奇广域透射电子显微镜作为许多人可用的高分辨率方法,并详细介绍了制备方案。

我们是一个从事体积 EM 的实验室,结构工作,要说血小板是如何激活的,它们是如何聚集的?它们如何形成导致止血的血栓,以及在过度刺激的情况下如何导致闭塞性凝血。这样做的前提是,如果我们知道结构,我们就可以更好地靶向药物并带来更好的结果。

和许多研究人员一样,我们在研究中使用小鼠模型,我们都面临着确定我们在小鼠中以及我们在小鼠动脉和静脉中的发现如何延续到人类更大动脉和静脉中的机制的挑战。我们都希望小鼠和人类的原理是一样的。如果您以我们的工作并使用容量 EM 方法为例,我们所做的重大贡献是其中之一,即血小板不仅仅是自杀式炸弹。

它们保留颗粒。他们做事是为了响应信号。它们具有稳定状态,这些稳定状态是正常血栓的一部分,也是包容性凝块的一部分。

第二,我们带来了一种感觉,即形成的血栓确实存在结构,它不仅仅是一堆砖头,而是一个有序的结构,其中血小板根据其活化等进行排序。第三,我们带来了一种感觉,即人们应该能够从结构中应用和理解需要刺激什么才能获得好的结果,或者需要抑制什么才能产生保护人们免受不良结果影响的结果。我们在近毫米距离的近纳米尺度上收集序列图像。

我们将这些组合成一个蒙太奇图像。因此,我们可以在接近纳米到接近毫米的尺度上观察图像中的任何给定点,这足以将单个特征置于整个血栓内。研究人员现在有一个强大的工具,他们可以提出新的问题,凝血级联反应和细胞信号或药物如何在血栓的局部到全球水平上在空间上发挥作用?

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关键字:穿刺伤口 止血 血栓 血小板聚集 电子显微镜 样品制备 广域成像 血栓形成 血管损伤 固定 包埋 染色

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