Name: combined near-infrared fluorescent imaging and micro-computed tomography for directly visualizing cerebral thromboemboli
Uploaded: 2016-09-25T14:00:00
Description: Watch this Scientific Journal Video about Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli at JoVE.com

这项工作是由韩国医疗技术的R＆D项目，卫生部和福利（HI12C1847，HI12C0066）的支持，生物与医药科技发展计划（2010-0019862）和全球研究实验室（GRL）方案（NRF-2015K1A1A2028228）国家研究基金会，由韩国政府资助。

在这个协议证明了所有的动物程序已经审查并东国大学一山医院动物护理和使用委员会批准，并按照NIH的指南中列出的照顾和动物使用的原则和程序进行的。 1.外源性形成凝块的制备荧光标记标记（图1） <ol><li>在用3％异氟烷，用30％的氧气（1.5L /分钟100％的氧气）混合感应腔麻醉的小鼠。通过观察肌张力，并确认不存在脚趾捏反射确保麻醉深度足够。 </li><li>放置在俯卧位的无菌披盖的动?...

<ol>
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E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperacute+direct+thrombus+imaging+using+computed+tomography+and+gold+nanoparticles.">Hyperacute direct thrombus imaging using computed tomography and gold nanoparticles.</a> Ann Neurol. 73 (5), 617-625 (2013).</li><li>Kim, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Imaging+of+Cerebral+Thromboemboli+Using+Computed+Tomography+and+Fibrin-targeted+Gold+Nanoparticles.">Direct Imaging of Cerebral Thromboemboli Using Computed Tomography and Fibrin-targeted Gold Nanoparticles.</a> Theranostics. 5 (10), 1098-1114 (2015).</li><li>Kim, D. 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我们展示了使用两个互补的分子成像技术用于在栓塞性中风的实验模型直接血栓成像：血纤维蛋白靶向金纳米颗粒（FIB-GC-金纳米粒子），用于在体内基于Micro-成像和一个FXIIIA针对前光学成像探针体内荧光成像。 FIB-GC-金纳米粒子的静脉内给药后，血栓成为可见的CT为致密的结构，后浓度的梯度引起的颗粒变得由纤维蛋白 - 靶向肽的作用（该颗粒的表面上）...

在一个6人将在其一生某个时候中风。缺血性中风是迄今为止最常见的中风类型，占所有中风病例的80％。因为血栓引起广大这些缺血性中风，存在直接血栓成像的兴趣日益增加。 据估计，约200万脑细胞大脑中动脉闭塞1的每一分钟期间死亡，导致的口号“时间就是大脑”。计算机断层扫描（CT）的研究可以快速完成，并且被广泛使用;因为这个原因，CT仍首选初始诊断和治疗超急性缺血性中风的成像。 CT是用于通知重要的早期决定特别有价值的：溶栓施用组织纤维蛋白溶酶原激活剂（tPA）和/或归类（triaging）到血管内凝块检索2。当前的CT-基于血栓成像，但是，不能连续跟踪大脑体内升血栓栓塞，因为它使用间接方法来证明血栓：由碘化造影血池的混浊后，血栓被表现为在容器填充的缺陷。有与碘造影剂的重复给药相关的剂量限值和风险，排除重复的血栓成像用这种方式。 因此，为直接成像方法在中风患者脑血栓迫切需要，允许更快和更好的治疗决定作出。我们提出通过增强的CT，目前使用的前线成像模态对行程的值，与使用血栓寻求纳米粒分子成像剂来实现这一点。 我们已经利用显微计算机断层摄影术（显微），高分辨率的离体或在体内 （小动物）的CT成像版本，允许快速的数据采集&lt;展示了使用本剂的SUP&gt; 3,4。即使有可供小动物显微（比从人类大小的扫描仪差很多）相对较差的软组织对比度，成像剂是能够寻求和使他们对CT，通过分子加强了“密集的血管征”高密度标记血栓成像。 补充CT技术，我们集团之前开发利用经Cy5.5近红外荧光（NIRF）探针可视化脑血栓负担5光学直接血栓成像技术。这是验尸大脑的体外技术，但高度敏感，并提供在研究环境体内数据确认。 拥有CT和NIRF都基于血栓寻求成像技术使我们能够比较和对比这些技术来实现对血栓和血栓成像在缺血性中风的发展进程中的作用非常翔实的数据。 HERE，我们描述了在体内的显微和体外成像NIRF的组合技术的详细协议直接看到血栓栓塞的栓塞性中风的小鼠模型。这些简单的和鲁棒的方法是通过在治疗期间在体内使在血栓负荷/分布和动态血栓演化的表征体内评估准确迅速和定量方式推进我们血栓性疾病的理解是有用的，其次是提供体外数据作为体内成像结果的确认对照和参考标准。

<table border="0" cellpadding="0" cellspacing="0" width="588">
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	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:155px;">Machines</td>
			<td style="width:219px;"></td>
			<td style="width:119px;"></td>
			<td style="width:96px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">microCT</td>
			<td>NanoFocusRay, JeonJu, Korea</td>
			<td>NFR Polaris-G90</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">NIRF imaging system</td>
			<td>Roper-scientific,Tucson, AZ</td>
			<td>coolsnap-Ez</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Laser Doppler flowmeter</td>
			<td>Perimed, Stockholm, Sweden</td>
			<td>PeriFlux System 5000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Surgical microscope</td>
			<td>Leica Microsystems, Seoul, Korea</td>
			<td>EZ4HD</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:155px;">Inhalation anesthesia machine</td>
			<td>PerkinElmer, Massachusetts, USA</td>
			<td>XGI-8</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">NFR control</td>
			<td>NanoFocusRay, JeonJu, Korea</td>
			<td>NFR Polaris-G90</td>
			<td style="width:96px;">microCT control software</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:155px;">Lucion</td>
			<td>Infinitt, Seoul, Korea</td>
			<td>Lucion</td>
			<td style="width:96px;">3D render imaging software</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lab chart 7</td>
			<td>ADInstruments, Colorado, USA</td>
			<td>Lab chart 7</td>
			<td>rCBF</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Image J software</td>
			<td>Wanye Rasband, NIH, USA</td>
			<td>1.49d</td>
			<td>imaging analysis</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Devices/Instruments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Infusion pump</td>
			<td>Harvard, Massachusetts, USA</td>
			<td>pump 22(55-2226)</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Homeothermic blanket</td>
			<td>Panlab, Barcelona, Spain</td>
			<td>HB101</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Pocket cautery</td>
			<td>Daejong, Seoul, Korea</td>
			<td>DJE-39</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Brain matrice</td>
			<td>Ted pella, CA, USA</td>
			<td>15003</td>
			<td style="width:96px;">coronal section</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">PE-50 tubing</td>
			<td>Natsume, Tokyo, Japan</td>
			<td>SP-45(PE-50)</td>
			<td style="width:96px;">I.D. 0.58 mm O.D. 0.96 mm</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">PE-10 tubing</td>
			<td>Natsume, Tokyo, Japan</td>
			<td>SP-10(PE-10)</td>
			<td style="width:96px;">I.D. 0.28mm O.D. 0.61 mm</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:155px;">30 gauge needle</td>
			<td>sungshim-medical, Seoul, Korea</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Syringe</td>
			<td>CPL-medical, Ansan, Korea</td>
			<td></td>
			<td>1 &#38; 3 cc</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Gauze</td>
			<td>Panamedic, Cheonan, Korea</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tape</td>
			<td>Scotch, Seoul, Korea</td>
			<td>3M-810</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Micro forceps</td>
			<td>Fine Science Tools, Vancouver, Canada</td>
			<td>&#160;11253-27</td>
			<td>Dumont #L5</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Micro scissor</td>
			<td>Fine Science Tools, Vancouver, Canada</td>
			<td style="width:119px;">15000-03</td>
			<td>Vannas spring</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Scissor</td>
			<td>Fine Science Tools, Vancouver, Canada</td>
			<td style="width:119px;">14084-08</td>
			<td>8.5 cm</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Black silk suture</td>
			<td>Ailee, Busan, Korea</td>
			<td>SK6071, SK728</td>
			<td>6-0 and 7-0</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">meloxicam</td>
			<td>Yuhan, Seoul, Korea</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">vet ointment</td>
			<td>Novartis, Basel, Swiss</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">10% Povidone-iodine (betadine)</td>
			<td>Firson, Cheon-an, Korea</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FeCl3</td>
			<td>Sigma, Missouri, United States</td>
			<td>157740-5G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">TTC</td>
			<td>Amresco, Ohio, USA</td>
			<td>0765-100g</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Isoflurane</td>
			<td>Hana-Pham, Gyeonggi, Korea</td>
			<td>Ifran</td>
			<td>100 mL</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">PBS</td>
			<td>Welgene, Daegu, Korea</td>
			<td style="width:119px;">LB001-02</td>
			<td>500 mL</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Gold nanoparticles</td>
			<td></td>
			<td></td>
			<td>Synthesis</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">C15 optical agent</td>
			<td></td>
			<td></td>
			<td>Synthesis</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Tissue plasminogen activator</td>
			<td style="width:219px;">Boehringer Ingelheim, Biberach, Germany</td>
			<td style="width:119px;">rtPA(actilyse)</td>
			<td>20 mg</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal saline</td>
			<td>Daihan Pham, Seoul, Korea</td>
			<td>48N3AF3</td>
			<td>20 mL</td>
		</tr>
	</tbody>
</table>

combined near-infrared fluorescent imaging and micro-computed tomography for directly visualizing cerebral thromboemboli

直接血栓成像可视化血栓梗塞的根本原因。如果能够像血栓允许直接行程不是依靠间接测量的更好的调查，将是一个强有力的和强大的血管的研究工具。我们使用标签与分子成像血栓血栓标记的光学成像方法 - 该凝块的成熟过程中被共价连接至由活化的凝血因子的ⅩⅢa的血纤维蛋白交联的酶作用的血栓的纤维蛋白丝一经Cy5.5近红外线荧光（NIRF）探针。的微型计算机断层扫描（显微）基的方法是使用官能化以靶向凝块的主要成分血栓求金粒子（AuNPs）：纤维蛋白。本文描述了组合的体内显微和栓塞性中风的小鼠模型血栓栓塞的体外 NIRF成像的详细协议。我们表明， 在体内&lt;/ em&gt;的显微和纤维蛋白靶向乙二醇壳聚糖纳米金（FIB-GC-金纳米粒子）可同时用于原位血栓和脑栓塞血栓可视化。我们还描述了使用在体内基于Micro-直接血栓成像以串行监控组织纤溶酶原激活介导的溶栓治疗效果。最后成像会议之后，我们证明通过体外 NIRF成像的程度和残留血栓栓塞的在脑中的分布。最后，我们描述了显微及成像NIRF数据定量分析图像。直接血栓成像的组合技术允许血栓可视的两个独立的方法来进行比较：血栓有关的荧光信号对区域体外 NIRF成像与高密度显微血栓体内的体积。

基线显微图像， 在体内得到施用FIB-GC-金纳米粒子（10毫克/毫升，300微升）以栓塞性中风后1小时，明显地显现在MCA脑血栓后-远端颈内动脉ACA分叉区（ 图6 ）。后续显微成像显示，用生理盐水治疗血栓COW没有变化。然而，tPA的治疗显示COW血栓的逐渐溶解（ 图6中的蓝色箭头）。这一发现表明，显微血栓成像可以允许溶栓治疗监测。有一个在24?...

Watch this Scientific Journal Video about Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli at JoVE.com

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

本协议描述相结合近红外荧光（NIRF）成像和微计算机断层扫描（显微）的可视化脑血栓中的应用。这种技术允许的血栓负担和演化的量化。该NIRF成像技术可视化荧光标记在切除脑血栓，而显微技术，可视化使用金的纳米粒子的生活里面的动物血栓。

结合近红外荧光成像和微计算机断层扫描直接可视化脑血栓

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Research

JoVE Journal

Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

体内近红外荧光（NIRF）血管内分子成像炎性斑块，多式联运方针成像动脉粥样硬化

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

科研

医学

Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging

微型计算机断层扫描成像制备小鼠冠状动脉血管逆行灌注和灌装

Visualization of the vasculature is becoming increasingly important for understanding many different disease states. While several techniques exist for ...

Near Infrared Optical Projection Tomography for Assessments of &beta;-cell Mass Distribution in Diabetes Research

Near Infrared Optical Projection Tomography for Assessments of ÃƒÅ½Ã‚Â²-cell Mass Distribution in Diabetes Research

近红外光学投影层析成像技术在糖尿病研究β细胞质量分布的评估

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

药物输送到微型计算机断层扫描成像缺血后肢血管灌注与微滤渗透

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery

组织鬼怪模拟，以评估乳癌手术潜在的近红外荧光成像中的应用

Inaccuracies in intraoperative tumor localization and evaluation of surgical margin status result in suboptimal outcome of breast-conserving surgery ...

Assessment of Viability of Human Fat Injection into Nude Mice with Micro-Computed Tomography

评估人体脂肪注射生存能力的裸鼠与微型计算机断层扫描

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft ...

Using Micro-computed Tomography for the Assessment of Tumor Development and Follow-up of Response to Treatment in a Mouse Model of Lung Cancer

采用微计算机断层扫描对肿瘤发展的评估和后续反应来治疗肺癌的小鼠模型

Lung cancer is the most lethal cancer in the world. Intensive research is ongoing worldwide to identify new therapies for lung cancer. Several mouse ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

"一站式" 近红外光谱对骨骼肌神经血管的耦合、氧化能力及微血管功能的作用

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

How to Administer Near-Infrared Spectroscopy in Critically ill Neonates, Infants, and Children

如何管理危重新生儿、婴儿和儿童近红外光谱

Near infrared spectroscopy (NIRS) calculates regional tissue oxygenation (rSO2) using the different absorption spectra of oxygenated and deoxygenated ...

Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination

近红外光免疫疗法为小鼠模型的胸膜传播

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination ...