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物，并使用吸入面罩和2％异氟醚，用30％的氧气混合，保持麻醉下。通过执行无菌技术和无菌衣/口罩/手套/仪器下面的步骤。保持通过清洗和前一消毒用70％酒精实验区在无菌条件第二手续后。 </li><li>心脏穿刺后6收集约300〜1000微升动脉血。混合70微升的全血用30微升C15探针5（20微摩尔/ L的浓度），一个经Cy5.5荧光探针来活化因子XIII（FXIIIA）凝血酶的血纤维蛋白交联的活性敏感，要荧光标记凝块（ 图1A ）。使用3毫升注射器（23号针）到一个20cm长的聚乙烯管（PE-50，ID0.58毫米）注入该混合血液。聚乙烯管道必须被消毒（或认证的由制造商无菌）和血块必须在组织培养罩无菌制备。 </li><li>通过观察呼吸不足和心脏脉搏的检查动物的死亡。 </li><li>留在室温下将血液装管2小时，然后在4℃下22小时，并执行下列步骤，如先前报道7。 </li><li>切含血栓管进入1.5厘米长的小段。使用3毫升注射器填充有磷酸盐缓冲盐水（PBS），通过轻轻注入PBS成每片管的驱逐血栓到含PBS-6孔板中。用PBS（ 图1B）洗血栓三次。 </li><li>通过仔细地绘制洗涤血栓，同时避免气泡，用生理盐水填充1ml注射器用30用1.5厘米长的血栓装载有15厘米长的PE-10管（ID0.28毫米）的前端部被插入到该管的近端号针头。 </li><li>血栓加载PE-10管连接与修改，以具有锥形端部（ID 200微米）一个3厘米长的PE-50管（ID0.58毫米），这将被放置在大脑中动脉（MCA） - 大脑前在栓塞性中风的小鼠模型（ 图1C）颈内动脉（ICA）的动脉（ACA）的分叉区域。 </li></ol> 2.造型血栓栓塞性中风的小鼠模型（图2） <ol><li> Anesthetiz如先前在用3％异氟烷，用30％的氧气（1.5升/分钟）混合感应室报告7被抚摸EA不同的鼠标。美洛昔康注射（5毫克/公斤）皮下注射，以减轻术后疼痛。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>适用兽医软膏少量每只眼睛，以防止麻醉期间干燥。通过执行无菌技术和无菌衣/口罩/手套/仪器下面的外科手术。保持通过在清洗之前，用70％的酒精消毒手术区，无菌条件下，并在手术后。 </li><li>将鼠标移动到手术台。放置在俯卧位的无菌披盖的动物，并使用吸入面罩和2％异氟烷保持麻醉下。然后，使用具有反馈的恒温毯从直肠温度计夹住体温在36.5℃。 </li><li> surgi后校准制备具有优碘和70％的酒精，通过使用在左耳和眼睛之间的头皮手术刀使1cm的垂直切口。胶光纤激光多普勒流量计到暴露的左颞骨表面的远端（1 Mm左和来自前囟低于4毫米）。然后，开始多普勒血流监测（ 图2A）。 </li><li>放下动物。通过拉动与附连到销的字符串上门牙拉直颈部和剃颈部区域。然后，使3厘米的垂直正中切口，传播它开放，通过解剖周围的血管软组织暴露左侧颈动脉球区。要小心，不要伤害迷走神经。 </li><li>结扎用消毒6-0黑丝线缝合左侧近端颈总动脉（CCA），以及结扎左ICA和使用无菌7-0黑丝线缝合左侧翼腭动脉（PPA）。 </li><li>烧灼左甲状腺动脉，这是左外颈动脉USI的一个分支NG单极电烧灼，结扎和左近端颈外动脉（ECA）松散和紧密用无菌7-0黑丝线缝合更远端站点。 </li><li>用微型剪刀，进行ECA的连接点之间的小孔（直径约0.2〜0.25微米）。 </li><li>将含血栓导管插入在ECA孔同时放松近端结扎ECA。推进导管进入CCA为了以收紧导管到位后，重新拧紧近端结扎ECA。 </li><li>烧灼切割ECA远端部分，也就是远离远端结扎部位，并旋转自由近端ECA顺时针它对齐到ICA的方向同时取出从CCA导管。然后，将导管大约9mm到MCA - 远端ICA的ACA分支区域松开ICA结扎后立即生效。然后，拧紧结扎ICA。 </li><li>放置血栓进入分叉区轻轻按压注射器1毫升注射血栓。检查多普勒脑血流量（CBF），这应该由30％或更低的降低与基线相比，如果血栓成功闭塞血管（ 图2B）的下降。 </li><li>取出导管，并立即紧紧地结扎ECA近端部位。此外，unligate共同国家评估和PPA。 </li><li>关闭用6-0丝线缝合切口部位。继续进行多普勒监测过的所需时间跨度（在此，为血栓注射后30分钟）后，停止麻醉。返回鼠标在一个空笼子，并保持温暖，取暖灯。直到它已获得了足够的意识，以保持胸骨斜卧不要离开鼠标无人值守。 </li></ol> 3. 在脑血栓的体内显微成像（图3） <ol><li>重新麻醉作为在预先指定的时间点在步骤2.1中所述，用2％异氟醚的小鼠（在此，1小时）的发作以下栓塞性中风的因血栓，在脑动脉的位置。通过确认不存在脚趾捏反射确保麻醉足够深度。适用兽医软膏少量每只眼睛，以防止麻醉期间干燥。 </li><li>在在10mM PBS中的10毫克的Au / ml的浓度重新悬浮的血纤维蛋白靶向金纳米颗粒（FIB-GC-金纳米粒子4），超声处理的血栓成像剂为30分钟，以确保纳米粒子的溶解和分散。注入300μl的FIB-GC-金纳米粒子（10毫克/毫升）插入阴茎静脉。 </li><li>放置在显微机的床中的动物，并且通过拉动与连接到一个销，以减少运动伪影的字符串上门牙拉直颈部。 </li><li>在FIB-GC-金纳米粒子的注射后5分钟，开始获取脑的显微图像。对于这里所描述的实验，请使用以下的成像参数:65 kVp的，60μA，观点26.7点¯x26.7点¯x27.9毫米3场，0.053点¯x0.053×0.054毫米3像素大小，每帧100毫秒，1平均，360视图，512×512的重建矩阵，600片，64秒的扫描时间。 </li><li>返回鼠标在一个空笼子，并保持温暖，取暖灯。直到它已获得了足够的意识，以保持胸骨斜卧不要离开鼠标无人值守。 </li><li>使用安装在显微扫描仪的一个软件包的" 重建 "面板的" 开始 "命令变换医学（DICOM）格式的原始图像数据转换成数字成像和通信。 </li><li>为图像（在步骤6）的定量分析，通过根据制造商的说明使用市售软件包变换DICOM数据转换成TIFF格式。 </li><li>定性分析和定量在一个更简单和更快速的方式进行分析，使用的软件包和原始0.054毫米厚图像以DICOM格式，用于产生一组新的呈现具有1或2毫米（这里，2mm）的厚度，如下轴向和冠状图像。 <ol><li>选择在" 数据源 "树中的DICOM文件夹，单击鼠标右键，并将该文件夹导入到"MasterDB&#39;或&#39;PrivateDB"。 </li><li>点击"MasterDB&#39;或&#39;PrivateDB"中的"数据源"树，然后选择导入文件夹。点击最左边的面板中的"3D"选项卡后，当"加载选项"窗口弹出时，按下" 确定 "导入图像序列中的文件夹作为一个堆栈英寸</li><li>通过点击在轴向和冠状图像窗口单词&#39; 的MRP&#39;和选择弹出菜单上&#39;MIP&#39;改变图像表示以最大强度投影（MIP）格式。在同一个窗口:然后，点击&#39;0毫米] TH&#39;后的图像厚度更改为2毫米。 </li><li>使用2mm宽的3D娜vigator栏，允许探索的图像堆栈，并在适当的角度和位置，它切片，与威利斯（COW）的圆形，其中港口血栓的全覆盖制备2mm厚的轴向截面图像。点击" 输出 "面板上的拍摄按钮（相机图标）。保存为TIFF格式的图像。 </li></ol></li><li>然后，准备五个双毫米，而且从额叶小脑连续覆盖厚冠状切面图像。 <ol><li> ACA血栓 - 首先，由轴向图像上仔细对齐导航栏，以便日冕图像最能直观马华准备第二片。 </li><li>接着，在连续方式制备其他四片。点击" 输出 "面板上的拍摄按钮（相机图标）。 TIFF格式保存图像。 </li></ol></li></ol> 4.溶栓及在脑血栓的体内显微成像（图3） <ol><li>准备100厘米长的PE-10管与在一端上有30号针头，而在另一端的1ml注射器。填用盐水（600微升）或tPA的（在此，24毫克/千克，600微升），同时避免气泡管。 </li><li>如在步骤2.1中所述重新麻醉小鼠。插入动物的阴茎静脉内针尖。小心地将动物到显微机的床。然后，固定并通过将其编带至床稳定导管系统的静脉内注射的一部分。 </li><li>执行后续成像会议前处理基线。然后，通过按压针筒柱塞到导管系统注入或60微升生理盐水或tPA的。在推注后，开始注入在一段时间（在此为30分钟）余下的溶液（540微升）。 </li><li>在预先指定的时间点获得使用相同的参数如在步骤3.4显微图像:在这里，在推注后3和24小时。要执行以下体外血栓NIRF大脑的总线成像，通过颈脱位安乐死麻醉下的动物。 </li></ol> 5. 离体 NIRF血栓影像与三苯四唑氯化物（TTC）的脑组织的染色（图4） <ol><li>断头后，取出头皮并通过与从枕骨大孔达向矢状缝剪刀剪头骨。通过仔细地提升用剪刀，同时避免伤害到下面的大脑颅骨切开的边缘，从而一语道破半球删除颅腔。 </li><li>切出视神经在大脑基底尽可能接近到脑表面，因为它们可以与威利斯动脉的圆，应该在NIRF成像可视化重叠。然后，为了清楚地可视化的Y形"MCA / ACA /远端ICA&#39;为NIRF血栓成像，之后轻轻压缩小脑基，以暴露第切出的远端的ICAs尽可能给脑表面Ë切割点（ 图4A）。 </li><li>执行NIRF成像（激发/发射675 / 690nm下1秒的曝光）及其碱朝上（ 图4B，C），其中可视化的荧光标记的血栓在COW的动脉去除的脑组织（ 图4D） 。然后，用脑朝上，该可视化皮质血栓栓塞的顶点执行附加成像。通过将盐水几滴在组织避免大脑的干燥。 </li><li>使用根据制造商的说明脑矩阵器件，快速地准备冠2mm厚的脑切片:6片2mm厚片（2.3，0.3，-1.7，-3.7，从前囟-5.7毫米）。通过使用盐水液滴避免大脑切片干燥，并执行前和各部分的背面两者的NIRF成像。 </li><li>放入2％三苯基氯化四氮唑（TTC）溶液中的大脑切片20分钟，同时避免暴露到二极管灯吨。然后，将切片在4％甲醛溶液在4℃下，同时避免暴露于光。 </li></ol>采用显微图像和ImageJ的（1.49d）软件6.血栓面积的定量（图5） <ol><li>打开图像（ 图5A）。 <ol><li>打开图像序列文件选择" 文件&gt;导入&gt;图像序列 "或" 文件&gt;打开 "创建堆栈文件。通过使用" 图像&gt;类型&gt; 8位 "将图像转换为8位灰度。 </li></ol></li><li>转换英寸该股毫米（ 图5A）。 <ol><li>当CT图像文件中的一个像素对应于0.053毫米，可以使用"分析&gt;设置缩放&#39;进入&#39;1&#39;，&#39;0.053"，并为" 距离像素 &#39;毫米&#39;，&#39; 已知距离 "，和长度的 &#39; 单元 &#39;， 分别。 </li><li>当（只）比例尺是可用的，使用"直线"工具绘制与它的长度等于标尺条的一条线。然后，使用" 分析&gt;设置规模 "，进入毫米的比例尺的长度。 </li></ol></li><li>背景减法（ 图5B） <ol><li>通过使用" 编辑&gt;选项&gt;指定&#39;的地方对脑实质的领域的兴趣（ROI）圆形或矩形区域无血栓或骨相关的高密度地区。因为指定的投资回报率适用于堆栈的每一个切片检查，以确保如果没有在每个片高密度的区域重叠。 </li><li>选择"插件&gt;投资回报率&gt;从投资回报率BG减法&#39;，为&#39;从平均标准偏差数"进入2.0。 </li></ol></li><li>血栓栓塞相关病变高密度分割（ 图5C） <ol><li>选择" 图像&gt;调整&gt;阈值"，然后输入罗"的价值观WER门限电平"和"上限程度"分别为22和255。选择"上/下"，以显示在下方的蓝色灰度，阈值像素下限阈值像素，并且像素绿色上限阈值以上。 </li><li>使用"写意选择工具"绘制围绕血栓相关的高密度灶，而不包括骨性领域的投资回报。在绘图过程中，按住要么[SHIFT]或[ALT]添加或删除一个区域，分别为。 注意:如果高密度血栓栓塞病灶从骨结构遥远，而不是&#39;手绘选择工具"，"棒工具"可以安全地在不改变它的"容差"电平使用。 </li></ol></li><li>该分段病变的定量分析（ 图5D） <ol><li>使用"分析&gt;设置测量&#39;选择&#39;区&#39;，&#39;平均灰度值"和"集成密度（区X平均值）9 ;.检查下列选项:"限价至阈值"和"显示标签"。使用"分析&gt;测量"来获取量化数据。然后，将结果保存为一个"的.xls"文件。 </li><li> TIFF格式存储堆栈文件。此外，使用" 分析&gt;工具&gt;投资回报率经理保存的ROI。 </li></ol></li></ol>

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catalytic+life+of+activated+factor+XIII+in+thrombi.+Implications+for+fibrinolytic+resistance+and+thrombus+aging.">Catalytic life of activated factor XIII in thrombi. Implications for fibrinolytic resistance and thrombus aging.</a> Circulation. 102 (10), 1151-1157 (2000).</li><li>Reed, G. L., Houng, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+contribution+of+activated+factor+XIII+to+fibrinolytic+resistance+in+experimental+pulmonary+embolism.">The contribution of activated factor XIII to fibrinolytic resistance in experimental pulmonary embolism.</a> Circulation. 99 (2), 299-304 (1999).</li><li>Sun, I. C., 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=Biocompatible+glycol+chitosan-coated+gold+nanoparticles+for+tumor-targeting+CT+imaging.">Biocompatible glycol chitosan-coated gold nanoparticles for tumor-targeting CT imaging.</a> Pharm Res. 31 (6), 1418-1425 (2014).</li><li>Celi, A., 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=Thrombus+formation:+direct+real-time+observation+and+digital+analysis+of+thrombus+assembly+in+a+living+mouse+by+confocal+and+widefield+intravital+microscopy.">Thrombus formation: direct real-time observation and digital analysis of thrombus assembly in a living mouse by confocal and widefield intravital microscopy.</a> J Thromb Haemost. 1 (1), 60-68 (2003).</li><li>Chen, I. Y., Wu, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+molecular+imaging:+focus+on+clinical+translation.">Cardiovascular molecular imaging: focus on clinical translation.</a> Circulation. 123 (4), 425-443 (2011).</li><li>Wintermark, M., 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=Imaging+recommendations+for+acute+stroke+and+transient+ischemic+attack+patients:+a+joint+statement+by+the+American+Society+of+Neuroradiology,+the+American+College+of+Radiology+and+the+Society+of+NeuroInterventional+Surgery.">Imaging recommendations for acute stroke and transient ischemic attack patients: a joint statement by the American Society of Neuroradiology, the American College of Radiology and the Society of NeuroInterventional Surgery.</a> J Am Coll Radiol. 10 (11), 828-832 (2013).</li><li>Weissleder, R., Tung, C. H., Mahmood, U., Bogdanov, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+tumors+with+protease-activated+near-infrared+fluorescent+probes.">In vivo imaging of tumors with protease-activated near-infrared fluorescent probes.</a> Nat Biotechnol. 17 (4), 375-378 (1999).</li><li>Narayanan, S., 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=Biocompatible+magnetite/gold+nanohybrid+contrast+agents+via+green+chemistry+for+MRI+and+CT+bioimaging.">Biocompatible magnetite/gold nanohybrid contrast agents via green chemistry for MRI and CT bioimaging.</a> ACS Appl Mater Interfaces. 4 (1), 251-260 (2012).</li><li>Amendola, V., 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=Magneto-plasmonic+Au-Fe+alloy+nanoparticles+designed+for+multimodal+SERS-MRI-CT+imaging.">Magneto-plasmonic Au-Fe alloy nanoparticles designed for multimodal SERS-MRI-CT imaging.</a> Small. 10 (12), 2476-2486 (2014).</li><li>Zhu, J., 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=Synthesis+of+Au-Fe3O4+heterostructured+nanoparticles+for+in+vivo+computed+tomography+and+magnetic+resonance+dual+model+imaging.">Synthesis of Au-Fe3O4 heterostructured nanoparticles for in vivo computed tomography and magnetic resonance dual model imaging.</a> Nanoscale. 6 (1), 199-202 (2014).</li></ol>

DE.K.，JY.K，通道A和KK是纤维蛋白靶向纳米金的专利持有人（10-1474063-0000，韩国知识产权局）。其余的作者都没有透露。

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

combined-near-infrared-fluorescent-imaging-micro-computed-tomography

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

9.8K 次观看.  Dongguk University College of Medicine. 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。 

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

Contrast Enhanced Vessel Imaging using MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between different soft tissues and body fluids is inherently poor in micro-CT images 1. Under these circumstances, visualization of blood vessels becomes a nearly impossible task. To overcome this and to improve the visualization of blood vessels exogenous contrast agents can be used. Herein, we present a methodology for visualizing the vascular network in a rodent model. By using a long-acting aqueous colloidal polydisperse iodinated blood-pool contrast agent, eXIA 160XL, we optimized image acquisition parameters and volume-rendering techniques for finding blood vessels in live animals. Our findings suggest that, to achieve a superior contrast between bone and soft tissue from vessel, multiple-frames (at least 5-8/ frames per view), and 360-720 views (for a full 360&deg; rotation) acquisitions were mandatory. We have also demonstrated the use of a two-dimensional transfer function (where voxel color and opacity was assigned in proportion to CT value and gradient magnitude), in visualizing the anatomy and highlighting the structure of interest, the blood vessel network. This promising work lays a foundation for the qualitative and quantitative assessment of anti-angiogenesis preclinical studies using transgenic or xenograft tumor-bearing mice.

Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s).

增强容器成像MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between ...

科研

医学

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

Visualization of the vasculature is becoming increasingly important for understanding many different disease states. While several techniques exist for imaging vasculature, few are able to visualize the vascular network as a whole while extending to a resolution that includes the smaller vessels1,2. Additionally, many vascular casting techniques destroy the surrounding tissue, preventing further analysis of the sample3-5. One method which circumvents these issues is micro-Computed Tomography (&mu;CT). &mu;CT imaging can scan at resolutions &lt;10 microns, is capable of producing 3D reconstructions of the vascular network, and leaves the tissue intact for subsequent analysis (e.g., histology and morphometry)6-11. However, imaging vessels by ex vivo &mu;CT methods requires that the vessels be filled with a radiopaque compound. As such, the accurate representation of vasculature produced by &mu;CT imaging is contingent upon reliable and complete filling of the vessels. In this protocol, we describe a technique for filling mouse coronary vessels in preparation for &mu;CT imaging.
Two predominate techniques exist for filling the coronary vasculature: in vivo via cannulation and retrograde perfusion of the aorta (or a branch off the aortic arch) 12-14, or ex vivo via a Langendorff perfusion system 15-17. Here we describe an in vivo aortic cannulation method which has been specifically designed to ensure filling of all vessels. We use a low viscosity radiopaque compound called Microfil which can perfuse through the smallest vessels to fill all the capillaries, as well as both the arterial and venous sides of the vascular network. Vessels are perfused with buffer using a pressurized perfusion system, and then filled with Microfil. To ensure that Microfil fills the small higher resistance vessels, we ligate the large branches emanating from the aorta, which diverts the Microfil into the coronaries. Once filling is complete, to prevent the elastic nature of cardiac tissue from squeezing Microfil out of some vessels, we ligate accessible major vascular exit points immediately after filling. Therefore, our technique is optimized for complete filling and maximum retention of the filling agent, enabling visualization of the complete coronary vascular network &#x2013; arteries, capillaries, and veins alike.

Visualization of the coronary vessels is critical to advancing our understanding of cardiovascular diseases. Here we describe a method for perfusing murine coronary vasculature with a radiopaque silicone rubber (Microfil), in preparation for micro-Computed Tomography (&mu;CT) 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 ...

Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

Small animal fluorescence molecular imaging (FMI) can be a powerful tool for preclinical drug discovery and development studies1. However, light absorption by tissue chromophores (e.g., hemoglobin, water, lipids, melanin) typically limits optical signal propagation through thicknesses larger than a few millimeters2. Compared to other visible wavelengths, tissue absorption for red and near-infrared (near-IR) light absorption dramatically decreases and non-elastic scattering becomes the dominant light-tissue interaction mechanism. The relatively recent development of fluorescent agents that absorb and emit light in the near-IR range (600-1000 nm), has driven the development of imaging systems and light propagation models that can achieve whole body three-dimensional imaging in small animals3.
Despite great strides in this area, the ill-posed nature of diffuse fluorescence tomography remains a significant problem for the stability, contrast recovery and spatial resolution of image reconstruction techniques and the optimal approach to FMI in small animals has yet to be agreed on. The majority of research groups have invested in charge-coupled device (CCD)-based systems that provide abundant tissue-sampling but suboptimal sensitivity4-9, while our group and a few others10-13 have pursued systems based on very high sensitivity detectors, that at this time allow dense tissue sampling to be achieved only at the cost of low imaging throughput. Here we demonstrate the methodology for applying single-photon detection technology in a fluorescence tomography system to localize a cancerous brain lesion in a mouse model.
The fluorescence tomography (FT) system employed single photon counting using photomultiplier tubes (PMT) and information-rich time-domain light detection in a non-contact conformation11. This provides a simultaneous collection of transmitted excitation and emission light, and includes automatic fluorescence excitation exposure control14, laser referencing, and co-registration with a small animal computed tomography (microCT) system15. A nude mouse model was used for imaging. The animal was inoculated orthotopically with a human glioma cell line (U251) in the left cerebral hemisphere and imaged 2 weeks later. The tumor was made to fluoresce by injecting a fluorescent tracer, IRDye 800CW-EGF (LI-COR Biosciences, Lincoln, NE) targeted to epidermal growth factor receptor, a cell membrane protein known to be overexpressed in the U251 tumor line and many other cancers18. A second, untargeted fluorescent tracer, Alexa Fluor 647 (Life Technologies, Grand Island, NY) was also injected to account for non-receptor mediated effects on the uptake of the targeted tracers to provide a means of quantifying tracer binding and receptor availability/density27. A CT-guided, time-domain algorithm was used to reconstruct the location of both fluorescent tracers (i.e., the location of the tumor) in the mouse brain and their ability to localize the tumor was verified by contrast-enhanced magnetic resonance imaging.
Though demonstrated for fluorescence imaging in a glioma mouse model, the methodology presented in this video can be extended to different tumor models in various small animal models potentially up to the size of a rat17.

Diffuse fluorescence tomography offers a relatively low-cost and potentially high-throughout approach to preclinical in vivo tumor imaging. The methodology of optical data collection, calibration, and image reconstruction is presented for a computed tomography-guided non-contact time-domain system using fluorescent targeting of the tumor biomarker epidermal growth factor receptor in a mouse glioma model.

电脑断层导引的时域漫荧光体层摄影术在小动物的肿瘤生物标志物的本土化

Small animal fluorescence molecular imaging (FMI) can be a powerful tool for preclinical drug discovery and development studies1. However, light ...

A Multicenter MRI Protocol for the Evaluation and Quantification of Deep Vein Thrombosis

We evaluated a magnetic resonance venography (MRV) approach with gadofosveset to quantify total thrombus volume changes as the principal criterion for treatment efficacy&#160;in a multicenter randomized study comparing edoxaban monotherapy with a heparin/warfarin regimen for acute, symptomatic lower extremities deep vein thrombosis (DVT) treatment. We also used a direct thrombus imaging approach (DTHI, without the use of a contrast agent) to quantify fresh thrombus. We then sought to evaluate the reproducibility of the analysis methodology and applicability of using 3D magnetic resonance venography and direct thrombus imaging for the quantification of DVT in a multicenter trial setting. From 10 randomly selected subjects participating in the edoxaban Thrombus Reduction Imaging Study (eTRIS), total thrombus volume in the entire lower extremity deep venous system was quantified bilaterally. Subjects were imaged using 3D-T1W gradient echo sequences before (direct thrombus imaging, DTHI) and 5 min after injection of 0.03 mmol/kg of gadofosveset trisodium (magnetic resonance venography, MRV). The margins of the DVT on corresponding axial, curved multi-planar reformatted images were manually delineated by two observers to obtain volumetric measurements of the venous thrombi. MRV was used to compute total DVT volume, whereas DTHI was used to compute volume of fresh thrombus. Intra-class correlation (ICC) and Bland Altman analysis were performed to compare inter and intra-observer variability of the analysis. The ICC for inter and intra-observer variability was excellent (0.99 and 0.98, p &#60;0.001, respectively) with no bias on Bland-Altman analysis for MRV images. For DTHI images, the results were slightly lower (ICC = 0.88 and 0.95 respectively, p &#60;0.001), with bias for inter-observer results on Bland-Altman plots. This study showed feasibility of thrombus volume estimation in DVT using MRV with gadofosveset trisodium, with good intra- and inter-observer reproducibility in a multicenter setting.

The goal of this study is to use magnetic resonance venography with long-circulating gadolinium-based contrast agent and direct thrombus imaging for quantitative evaluation of DVT volume in a multicenter, clinical trial setting. Inter- and intra-observer variability assessments were conducted, and reproducibility of the protocol was determined.

深静脉血栓形成的评价和定量的多中心MRI协议

We evaluated a magnetic resonance venography (MRV) approach with gadofosveset to quantify total thrombus volume changes as the principal criterion for ...

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel. 
The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke.

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo:  A Model of Single Vessel Stroke

Here, we describe a semi-invasive optical microscopy approach for the induction of a Rose Bengal photothrombotic clot in the somatosensory cortex of a mouse in vivo. The technical aspects of the imaging procedure are described from induction of a photothrombotic event to application and data collection.

玫瑰红Photothrombosis共聚焦光学成像<em&gt;在体内</em&gt;:单支脑卒中模型

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to ...

Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles has emerged as an effective method to transiently and locally increase the permeability of the BBB, facilitating para- and transcellular transport of drugs across the BBB. Imaging the vasculature during ultrasound-microbubble treatment will provide valuable and novel insights on the mechanisms and dynamics of ultrasound-microbubble treatments in the brain.
Here, we present an experimental procedure for intravital multiphoton microscopy using a cranial window aligned with a ring transducer and a 20x objective lens. This set-up enables high spatial and temporal resolution imaging of the brain during ultrasound-microbubble treatments. Optical access to the brain is obtained via an open-skull cranial window. Briefly, a 3-4 mm diameter piece of the skull is removed, and the exposed area of the brain is sealed with a glass coverslip. A 0.82 MHz ring transducer, which is attached to a second glass coverslip, is mounted on top. Agarose (1% w/v) is used between the coverslip of the transducer and the coverslip covering the cranial window to prevent air bubbles, which impede ultrasound propagation. When sterile surgery procedures and anti-inflammatory measures are taken, ultrasound-microbubble treatments and imaging sessions can be performed repeatedly over several weeks. Fluorescent dextran conjugates are injected intravenously to visualize the vasculature and quantify ultrasound-microbubble induced effects (e.g., leakage kinetics, vascular changes). This paper describes the cranial window placement, ring transducer placement, imaging procedure, common troubleshooting steps, as well as advantages and limitations of the method.

This protocol describes the surgical and technical procedures that enable real-time in vivo multiphoton fluorescence imaging of the rodent brain during focused ultrasound and microbubble treatments to increase blood-brain barrier permeability.

实时活体内多光子显微镜可可视化聚焦超声和微气泡处理，以提高血脑屏障通透性

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles ...

生物工程

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor determining patient outcome and is therefore frequently taken as a study endpoint. However, in small animal studies on SAH, quantification of cerebral vasospasm is a major challenge. Here, an ex vivo method is presented that allows quantification of volumes of entire vessel segments, which can be used as an objective measure to quantify cerebral vasospasm. In a first step, endovascular casting of the cerebral vasculature is performed using a radiopaque casting agent. Then, cross-sectional imaging data are acquired by micro computed tomography. The final step involves 3-dimensional reconstruction of the virtual vascular tree, followed by an algorithm to calculate center lines and volumes of the selected vessel segments. The method resulted in a highly accurate virtual reconstruction of the cerebrovascular tree shown by a diameter-based comparison of anatomical samples with their virtual reconstructions. Compared with vessel diameters alone, the vessel volumes highlight the differences between vasospastic and non-vasospastic vessels shown in a series of SAH and sham-operated mice.

The goal of this article is to present a method that allows a 3-dimensional reconstruction of the cerebrovascular tree in mice after micro computed tomography and determination of volumes of entire vessel segments that can be used to quantify cerebral vasospasm in murine models of subarachnoid hemorrhage.

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

神经科学

In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy

Cerebrospinal fluid (CSF) flow in rodents has largely been studied using ex vivo quantification of tracers. Techniques such as two-photon microscopy and magnetic resonance imaging (MRI) have enabled in vivo quantification of CSF flow but they are limited by reduced imaging volumes and low spatial resolution, respectively. Recent work has found that CSF enters the brain parenchyma through a network of perivascular spaces surrounding the pial and penetrating arteries of the rodent cortex. This perivascular entry of CSF is a primary driver of the glymphatic system, a pathway implicated in the clearance of toxic metabolic solutes (e.g., amyloid-&#946;). Here, we illustrate a new macroscopic imaging technique that allows real-time, mesoscopic imaging of fluorescent CSF tracers through the intact skull of live mice. This minimally-invasive method facilitates a multitude of experimental designs and enables single or repeated testing of CSF dynamics. Macroscopes have high spatial and temporal resolution and their large gantry and working distance allow for imaging while performing tasks on behavioral devices. This imaging approach has been validated using two-photon imaging and fluorescence measurements obtained from this technique strongly correlate with ex vivo fluorescence and quantification of radio-labeled tracers. In this protocol, we describe how transcranial macroscopic imaging can be used to evaluate glymphatic transport in live mice, offering an accessible alternative to more costly imaging modalities.

Transcranial optical imaging allows wide-field imaging of cerebrospinal fluid transport in the cortex of live mice through an intact skull.

使用荧光宏镜检查,通过"完整小鼠头骨"进行脑脊液传输的 Vivo 成像

Cerebrospinal fluid (CSF) flow in rodents has largely been studied using ex vivo quantification of tracers. Techniques such as two-photon microscopy and ...

Quantification of Cerebral Perfusion using Laser Speckle Imaging and Infarct Volume using MRI in a Pre-clinical Model of Posterior Circulation Stroke

Background: Basilar artery occlusion (BAO) is a subset of posterior circulation stroke that carries a mortality as high as 90%.&#160; The current clinical standard to diagnose ischemic stroke include computerized tomography (CT), CT angiography and perfusion and magnetic resonance imaging (MRI). Large animal pre-clinical models to accurately reflect the clinical disease as well as methods to assess stroke burden and evaluate treatments are lacking.
Methods: We describe a canine model of large vessel occlusion (LVO) stroke in the posterior circulation, and developed a laser speckle imaging (LSI) protocol to monitor perfusion changes in real time.&#160; We then utilized high b-value DWI (b=1800s/mm2) MRI to increase detection sensitivity. We also evaluated the ability of magnetic resonance angiography (MRA) to assess arterial occlusion and correlate with DSA. Finally, we verified infarct size from apparent diffusion coefficient (ADC) mapping with histology.&#160;
Results:&#160; Administration of thromboembolism occluded the basilar artery as tracked by DSA (n=7).&#160; &#160;LSI correlated with DSA, demonstrating a reduction in perfusion after stroke onset that persisted throughout the experiment, allowing us to monitor perfusion in real time.&#160; DWI with an optimized b-value for dogs illustrated the stroke volume and allowed us to derive ADC and magnetic resonance angiography (MRA) images. The MRA performed at the end of the experiment correlated with DSA performed after occlusion. Finally, stroke burden on MRI correlated with histology.
Conclusions: Our studies demonstrate real time perfusion imaging using LSI of a canine thromboembolic LVO model of posterior circulation stroke, which utilizes multimodal imaging important in the diagnosis and treatment of ischemic stroke.

A canine model of LVO stroke was utilized to develop laser speckle imaging to monitor cerebral perfusion in real-time.&#160; Diffusion-weighted MRI was optimized to image infarct volume utilizing a high b-value, enabling ADC and MRA, correlated with DSA at the time of stroke.&#160; Finally, ADC reconstructions correlated with histological findings.

Background: Basilar artery occlusion (BAO) is a subset of posterior circulation stroke that carries a mortality as high as 90%.&#160; The current clinical ...

蛛网膜下腔出血后脑周细胞收缩力的可视化

Imaging Vital and Non-vital Brain Pericytes in Brain Slices following Subarachnoid Hemorrhage

Pericytes are crucial mural cells situated within cerebral microcirculation, pivotal in actively modulating cerebral blood flow via contractility adjustments. Conventionally, their contractility is gauged by observing morphological shifts and nearby capillary diameter changes under specific circumstances. Yet, post-tissue fixation, evaluating vitality and ensuing pericyte contractility of imaged brain pericytes becomes compromised. Similarly, genetically labeling brain pericytes falls short in distinguishing between viable and non-viable pericytes, particularly in neurologic conditions like subarachnoid hemorrhage (SAH), where our preliminary investigation validates brain pericyte demise. A reliable protocol has been devised to surmount these constraints, enabling simultaneous fluorescent tagging of both functional and non-functional brain pericytes in brain sections. This labeling method allows high-resolution confocal microscope visualization, concurrently marking the brain slice microvasculature. This innovative protocol offers a means to appraise brain pericyte contractility, its impact on capillary diameter, and pericyte structure. Investigating brain pericyte contractility within the SAH context yields insightful comprehension of its effects on cerebral microcirculation.

作者聚焦:啮齿动物蛛网膜下腔出血后周细胞成像 

The preliminary inquiry confirms that subarachnoid hemorrhage (SAH) causes brain pericyte demise. Evaluating pericyte contractility post-SAH requires differentiation between viable and non-viable brain pericytes. Hence, a procedure has been developed to label viable and non-viable brain pericytes concurrently in brain sections, facilitating observation using a high-resolution confocal microscope.

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

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