伦理学声明：所有实验均批准的机构动物护理和使用委员会。根据机构指引照顾动物了。 ICR雌性小鼠（7 - 8周龄，25 - 30克）被安置在了空调，光控动物特拉维夫大学医学系赛克勒设施。在术语，动物安乐死麻醉过量。 1. Fibred共聚焦荧光显微镜（FCFM）校准<ol><li>打开该设备。 </li><li>连接探针（mini0 / 30）。 </li><li>根据制造商的说明校准装置。 </li></ol> 2.准备小鼠成像<ol><li>通过皮下注射既Ketaset（100毫克/千克）和XYL-M2（6毫克/千克）的麻醉。确定合适的麻醉通过反应迟钝到脚趾捏。 </li><li>切开腹股沟皮肤下面以<html>揭示了股动脉血管。请跟随切口切口部位湿润用生理盐水。 </li><li>用袋（或手套）约30秒盛有温水（不烫手）加热尾巴。制备的静脉内（IV）的分流为FITC葡聚糖（造影剂）的施用与盐水或化疗剂，通过插入一针（30克，1/2英寸）至尾静脉，并附加一个1ml注射器将其。确保静脉注射生理盐水打开。 注：FITC葡聚糖的IV施用（高分子量; 100微升; 10毫克/毫升; 2000 kDa的）便于可视化股骨微血管由FCFM。阿霉素（100微升; 8毫克/公斤，阿霉素），或盐水也将在后面IV施用到预加热的尾静脉。 </li><li>仰起将鼠标上的聚苯乙烯阶段。确保鼠标垫，用手术胶带保持位置。 </li></ol>标题“&gt; 3。显像股血管通过FCFM在和阿霉素或生理盐水给药后注意：在此研究中使用的纤维质共聚焦显微镜包括两个单元：（1）微探针（mini0 / 30）。 （2）激光扫描单元（LSU-488; 488nm的波长下）。 <ol><li>执行所有时间圈分析使用LSU 488纳米波长的激光。 注：主单元探测器探测过滤（500 - 650纳米）发出荧光。所获取的图像被事后重建，并在12帧/秒的速率显示。 </li><li>小心地断开从针的注射器，并附加包含FITC标记的葡聚糖的新注射器。管理（IV）100微升FITC右旋糖酐。 </li><li>移的探针（mini0 / 30），视合适字段和固定它，以下调整z轴，以获得相应的图像。等待开始信号淡出直到一个明确的和聚焦的信号是VISIBLE。 </li><li>记录基线血流短暂的稳定期（〜30秒）。然后连接到另一针注射器，要么含有阿霉素或生理盐水。四，管理100微升阿霉素或生理盐水。 </li><li>监视注入FITC-葡聚糖的流量连续地进行20分钟。在FCFM相关软件，使用直径按钮的上标尺以测量血管和它们分类为小（&lt;15微米）或大（&gt; 15微米）。 </li><li>安乐死麻醉过量的动物。 </li></ol>

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评价化疗诱导的血管毒性是具有挑战性，因为在可视化血管的动态响应于实时一个刺激的难度。大量临床研究已经牵连的几个化疗造成直接损伤血管，但这种毒性的机制和特点仍有待澄清。我们建立了实时， 体内分子成像的平台如本文8-10描述评价化疗在含有纤维状共聚焦荧光显微镜的小鼠的潜在血管毒性。这种高解析度的分子的小鼠成像是适合用于可视化动脉血流和血管&#39;架构?...

临床证据表明，几类化疗引起的各种由雷诺现象，高血压，心肌梗死，脑血管攻击，肝静脉occlusivedisease 1,2体现血管病症。 “&#39;意外&#39;抗血管生成药物”是一个相当新的名词，它描述了传统的化疗药物是作为可能的血管生成抑制剂，虽然他们不是最初开发用于这一目的3-5，但设计的征收尽可能少的消除肿瘤细胞的“抵押品损害“正常细胞尽可能3。几种化疗已经暗示为血管 - 毒物在应用血清生物标记物的临床研究中观察到。在这些被烷化剂（如环磷酰胺），铂化合物（例如顺铂）和蒽环类1,2,5-7。 急性心脑血管并发症，可能会出现一个resul吨血管毒性化疗所致的。他们可能会进步到像动脉粥样硬化，占后期血管发病的危险性增加慢性疾病。然而，尽管安装的临床证据，有指定的研究强调血管毒性的机制的缺乏，因此，它们造成的确切发病机理的进一步澄清是必要的。 在揭示化疗引起的血管毒性机制的一个主要挑战来源于体内调查血管功能的复杂性。我们描述了本文中的高清晰度的体内分子成像的小鼠，使捕捉血流和血管&#39;特性的一个平台。这个平台便于直接治疗性血管效应的检测：实时，以及跟随它们过了一段时间的同一个体中。

<table border="0" cellpadding="0" cellspacing="0" width="1395">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:579px;">Name of Material/ Equipment</td>
			<td style="width:423px;">Company</td>
			<td style="width:213px;">Catalog Number</td>
			<td style="width:183px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">general anesthesia (100mg/kg ketaset and 6mg/kg XYL-M2).&#160;</td>
			<td>Fort Dodge Animal Health, IA, USA and Biove Laboratories, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">a depilatory cream (Veet)</td>
			<td>&#160;ReckittBenckiser, Bristol, UK</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">A 30-gauge, 1/2-inch needle attached to 1 ml syringe&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FITC dextran at a volume of 100 &#181;l (10 mg/ml; FD2000S, MW 2000000 Dalton)&#160;</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Doxorubicin (8 mg/kg, Adriamycin)</td>
			<td>Teva, Israel</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">paclitaxel (1.2 mg/kg, Medexel)&#160;</td>
			<td>Taro, Israel</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Saline</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

chemotherapy-induced vascular toxicity - real-time in vivo imaging of vessel impairment

化学疗法的某些类可能施加急性血管变化，可能进展成长期条件可能会使病人血管并发症的风险增加。然而，尽管所述安装临床证据，有血管毒性，因此，血管/心血管病症一组异质性的病因的明确研究的缺乏尚待阐明。此外，该可依据血管毒性的机制可以完全由化疗引起的心脏毒性，这是直接细胞损伤相关的原理不同。我们建立了实时， 体内分子成像平台以评价抗癌疗法的潜在急性血管毒性。 我们已经建立了体内 ，小鼠的高分辨率分子成像，一个平台适用于密闭机关和参照一个BLO内血管可视化而每个单独的同一个体内外径船只充当其自身的控制。阿霉素给药后的血管壁均受损，代表血管毒性的一个独特的机制，可能是在终末器官损伤的早期事件。在此，纤维状共聚焦荧光显微镜（FCFM）基于成像的方法进行说明，它提供了一个创新的方式来了解生理现象在细胞和亚细胞水平的动物受试者。

体内连续成像在实时此处所使用的成像装置是一个高清晰度，纤维状共焦显微镜，装备有探针，使脉管的可视化和其响应于各种刺激如化疗。这种方法是微创因为尽管它可以便于深血管或器官的成像，它需要一个小切口的探针。探头束组成的纤维，光学显微镜和专有精密连接器数万。的示意图示于图1。 股骨头微血管成像<p class="jove_conte...

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Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

化疗引起的血管毒性 - 实时<em&gt;在体内</em&gt;船舶减值成像

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an ...

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Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

7.3K 次观看.  Tel Aviv University. We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.