This work was supported by the National Institutes of Health (HL114669). CM is a recipient of a postdoctoral research fellowship grant awarded by the Myocarditis Foundation, NJ.

操守准则： 所有动物的程序均按照实验动物的护理和使用的指导原则进行，并经内布拉斯加林肯，内布拉斯加州林肯大学。 1，诱导实验性自身免疫性心肌炎<ol><li>通过在1x磷酸盐缓冲盐水中，以2 mg/1.5 ml的终浓度溶解MYHC-α334-352制备肽溶液。 </li><li>加入1无菌的1×PBS中的溶液，以含50μg冻干的PT，得到储备液浓度为50纳克/微升的小瓶中制备的百日咳毒素（PT）。取20微升的股票进入无菌1.5mL管中，加入980微升无菌1X PBS中，获得1纳克/微升的工作浓度。 </li><li>通过添加40毫克结核分枝杆菌 H37Rv的制备完全弗氏佐剂（CFA）（MTB）提取至10ml CFA，得到的终浓度5毫克/毫升。 </li><li>准备肽终审法院乳液。注：对于EAM感应，肽终审法院乳液给予150微升含100微克MYHC-α每334-352动物。例如，免疫10小鼠应用1.5毫升含1毫克MYHC-α334-352的肽-CFA乳液。 <ol><li>为了制备1.5ml乳液，分装750微升的MYHC-α334-352肽溶液在一个1.5mL管中，和CFA辅以山地车到另一个1.5mL管中。使用3毫升鲁尔乐注射器，吸取上述肽溶液第一，其次是终审法院/ MTB提取物。 </li><li>连接注射器向3路活塞和活栓的另一出口连接到一个空3毫升注射器。调节活塞的通畅使肽混合物，终审法院从一个注射器流向其他与合理性好。 </li><li>通过反复按压的内容从一个针筒到另一为〜1分钟，然后将整个组件在冰上〜3分钟混合。重复此程序至少10倍。 </li><li>通过仔细地放置一个极小的液滴上的静水中的100毫升的玻璃烧杯中，测定该乳液的一致性。液滴预计不会分散在水中。如果这样做，继续混合，直至所需的一致性的实现。 </li><li>从3毫升注射器转移乳剂的内容到1毫升Luer-Lok接头的注射器通过更换附加到与1-ml注射器的活塞的两个3毫升的注射器中的一个，并附加一个27½号针头至1ml注射器的。 </li></ol></li><li>注入150μL肽终审法院乳液的分割剂量皮下注射在六到八周龄雌性的A / J小鼠（〜每75微升）两腹股沟区域。 </li><li>施用100μl的PT悬浮液（100毫微克）的腹腔内每只动物在第0天和第2天免疫后。 </li><li>通过施用150微升肽终审法院乳液的分割剂量皮下注射入T的两侧，重复第7天免疫程序他胸骨（〜75每微升）。准备这个乳液新鲜如上。然后，在第21天，若使动物MRM成像，见步骤3。 </li></ol> 2，动物处理<ol><li>将每个小鼠在含有2％异氟醚-空气混合物与加热垫置于下方，以保持温暖麻醉诱导室，以及动物转移到一个专门设计的动物固定器（ 图1）。 </li><li>固定化的动物中的动物支架俯卧位，以便其口鼻部配合到鼻锥维持麻醉（ 图1）。固定小鼠的头部与一个咬合杆连接到鼠标的门牙。 </li><li>打开空气吹加热器插在扫描仪的垂直孔中，以在实验过程中保持动物的体温其出口软管。 </li><li>维持麻醉以0.5〜2％异氟烷以2 ml / min的整个成像会话期间的流速。确认用脚趾掐法麻醉，没有动静预期。 </li><li>设置的充气枕头传感器，鼠尾/踝光纤脉搏血氧饱和度传感器，和直肠温度探头监控呼吸，心脏速率和体温，分别为（ 图1）。注：心脏门控是通过脉搏血氧饱和度，这使得无创监测动脉血氧饱和度进行。脉搏血氧饱和度传感器应安装在左侧踝关节和脚应该固定用螺纹环和录音保持在传感器的脚踝。 MRM成像是通过选通的呼吸和心脏信号，而无需使用任何造影剂来实现。 </li></ol> 3，图像采集<ol><li>制备动物（ 图1）后，将鼠标在MRM扫描仪的中心，定位在视场（FOV），其中磁场均匀性是最大的中心心脏。注：大口径（89毫米）9.4Ŧ垂直孔磁铁装有100克/厘米三重轴梯度和一个4厘米无线电频率（RF）的成像线圈，是用于获得高分辨率的三维（3D）图像。注意：一定要非磁性物料中使用核磁共振成像扫描仪的时候。 </li><li>运行成像接口，并从“学习选项”菜单中选择“新探”。输入“mtune中”，在命令栏并运行它拉上了“调整图形用户界面”。然后选择“开始探头调整”，并点击“自动缩放”按钮。该调整GUI将改为显示RFsignal。使用远程调谐/匹配旋钮在线圈调谐RF线圈的质子共振频率（400兆赫）的端部。在“开始”选项卡进入“预扫描”页面通过点击相关按钮来运行的频率和功率校准。打标签的“学习垫片”的标签“&#39;上的XYZ（快速）按钮拉起垫补页面。转到垫片页面，选择所有的迭代，并创下垫片按钮来进行自动匀场。 </li><li>从成像界面中的“研究”选项卡本地化的小鼠心脏中选择一个侦察兵序列。在“采集”选项卡改变视场为35 mm 2，保持机器的默认设置。单击开始运行序列;调节动物座的位置，如果心脏是不是在视场中心，重新调谐射频线圈，并再次获得了球探的图像。 </li><li>点击“分析”选项卡上的“宝石”序列按钮，然后输入相应的“采集”选项卡上的采集参数，以获得两个正交平面沿短轴和心脏20,21的长轴。注：典型的采集参数的梯度回波扫描是：切片厚度1毫米;重复时间（TR），200毫秒;回波时间（TE），2.67毫秒（最小）;翻转角，25°;面内矩阵大小，128×128; FOV，22毫米2;多项收购，4;和1分30秒的一个近似的采集时间。 </li><li>在“高级”选项卡中选中“CINE”序列收集脉搏血氧仪门控短轴电影MR图像以测量左室解剖和功能参数。调整成像片的基础上，心脏的鼠标悬停和拖拽长轴切面的位置和角度。进入“采集”选项卡上的成像参数，以获得梯度回波电影序列：TR，500毫秒; TE，5毫秒; FOV 22 平方毫米;采集矩阵256×256;切片厚度为1mm;多项收购，8;帧数，6;和〜17分钟的采集时间。单击开始开始收购。 </li><li>转换使用图像处理软件的“I / O”选项卡将采集的图像以DICOM格式和相应的文件传输到数据中心进行处理。 </li><li>在成像过程结束时，所有嗷嗷小鼠从麻醉中的过滤器顶笼子内恢复。不要让老鼠无人看管，直到他们重新获得足够的意识，保持胸骨斜卧，并根据需要让他们继续深造。 </li></ol> 4，数据的心脏核磁共振显微成像分析<ol><li>使用分部软件分析的LV 19的解剖和功能参数。通过加载DICOM格式的电影影像到软件中使用“打开图像文件（S）”中的“文件”菜单的子菜单。在GUI上选择“MRGE&#39;作为成像技术，”日比“作为图像类型和&#39;短轴中心”作为图像视图平面。 </li><li>指定的时间范围，通过用于舒张末期和收缩末期的“设置当前期限为舒张末期”和“设置当前期限为收缩末期”分别在“编辑”菜单的子菜单。 </li><li>首先单击“LV”命令按钮，然后单击“远藤”和右下角面板上的“盈”命令按钮手动分别绘制左心室心内膜和心外膜。通过按相应的命令按钮，以增加计算的精度除去乳头肌。 </li><li>阅读LV的量化参数，如左室舒张末期容积，收缩末期容积，每搏输出量，以及右上角的面板上射血分数。点击“杂项”命令按钮，然后单击“测量卡尺”命令按钮来测量左室参数，如壁厚和心室内径22。 </li><li>单击“文件”菜单中的“保存两个图像堆栈和分割”子菜单来保存图像，包括分割，在&#39;垫&#39;格式需要重新处理的图像。 </li></ol>

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No conflicts of interests were declared.

这项研究描述为一种非侵入性工具，以确定心脏异常的影响与自身免疫性心肌炎小鼠的MRM过程及其效用。由于EAM的组织学特征类似于人类感染后心肌炎小鼠模型通常用来描绘心肌损伤23-25 ​​的免疫机制。但是，患有心肌炎动物出现临床表现正常，诊断是基于在组织学实验7终止进行。动物通常是牺牲在21天接种后。评估这样的疾病过程在单个时间点的限制使用这些模型，尤其是在...

心脏衰竭是死亡的主要原因，心肌炎是心脏衰竭的青春少年1一个最主要的原因。患有心肌炎患者大多没有症状和疾病自发缓解2。然而，那些受影响的10-20％可发展为慢性疾病，导致扩张性心肌病（DCM）3。各种动物模型已经发展到研究心肌炎的免疫发病机制。这种疾病可以通过免疫动物与心肌肌球蛋白重链（MYHC）-α或其免疫显性肽片段或通过与病原体如柯萨奇病毒B3 4-9感染诱导的心肌炎敏感的A / J和Balb / c小鼠。这项研究涉及MYHC-α在A / J小鼠334-352引起的心肌炎。尽管显示心肌浸润，心肌炎影响的动物出现临床表现正常;诊断依据心脏炎症7的组织学评价ND超声心动图10。 磁共振显微镜（MRM）是一种常用的方法，得到心血管成像具有高分辨率的三维平面上，允许的功能细节，以评估微小血管的水平（高达10微米直径），但分辨能力的这种电平是无法实现与常规的磁共振成像（MRI）扫描程序，其中，该分辨率通常是获得可达1毫米11-14。 MRM提供了一个优点，因为它允许获得的高分辨率图像，以及在疾病过程14的早期时间点获得的性能参数。在临床上，MRM成像已被广泛应用于研究的病变心脏，肺或脑部15-17功能参数。在这项研究中，显示了一个使用MRM技术作为一种非侵入性工具，以确定受影响的自身免疫性心肌炎的A / J小鼠的心脏异常。具体来说，T他MRM成像允许的功能参数如左室（LV）舒张末期容积和射血分数（EF）以合理的准确性18定量。各参数的定义如下：LV舒张末期容积，血容量在左心室的舒张周期的结束，和射血分数，每搏输出量/舒张末期容积。使用对通过磁共振扫描器19获取的处理符合DICOM标准的心血管图像而开发的免费提供的分类软件进行数据分析。该数据显示在心肌炎动物的增加左心室壁的厚度，对应于左室舒张末期容积，每搏输出量和射血分数下降，与健康小鼠这些功能参数进行比较。

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			<td height="25" style="height:25px;width:509px;">Myhc-a 334-352 (DSAFDVLSFTAEEKAGVYK)</td>
			<td style="width:364px;">Neopeptide, Cambridge, MA</td>
			<td style="width:145px;"></td>
			<td style="width:167px;">Store at 4&#959;C</td>
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			<td style="width:364px;">Sigma Aldrich, St Louis, MO</td>
			<td style="width:145px;">5881</td>
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			<td>Difco Laboratories, Detroit, MI</td>
			<td>231141</td>
			<td>Store at 4&#959;C</td>
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			<td>List Biologicals Laboratories, Campbell, CA</td>
			<td>181</td>
			<td>Store at 4&#959;C</td>
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			<td>Corning, Manassas, VA</td>
			<td>21-040-CV</td>
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			<td>Piramal Healthcare, Mumbai, India</td>
			<td>NDC66794-013-25</td>
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			<td>Jackson Laboratories, Bar Harbor, ME</td>
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			<td>309657</td>
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			<td>BD, Franklin Lakes, NJ</td>
			<td>305195</td>
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			<td height="21" style="height:21px;">Sterile needle, 27 1/2 G</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>305109</td>
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			<td>Smiths Medical ASD, Inc. Dublin, OH</td>
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			<td>6720</td>
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			<td>34155</td>
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			<td>Medline Industries, Mundelein, IL</td>
			<td>30-1001</td>
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			<td>Theramics Corporation, Springfield, IL</td>
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			<td height="21" style="height:21px;">Reptile heating lamp&#160;</td>
			<td>Energy Savers Unlimited, Inc. Carson, CA</td>
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			<td>Target Corporation, MN</td>
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			<td height="21" style="height:21px;">Up and Up Polymyxin B sulfate/Bacitracin/Neomycin sulfate antibiotic ointment</td>
			<td>Target Corporation, MN</td>
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			<td height="21" style="height:21px;">North Safety DeciDamp-2PVC foam ear plugs</td>
			<td style="width:364px;">North Safety Products, Smithfield, RI</td>
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			<td height="21" style="height:21px;">Cotton tipped applicator, 6&#8217;&#8217; wooden stem&#160;</td>
			<td>Jorgensen Laboratories, Inc. Loveland, CO</td>
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			<td>Summit Anesthesia Solutions, Ann Harbor, MI</td>
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			<td height="21" style="height:21px;">Summit Anesthesia Support system for regulating flow of anesthesia&#160;</td>
			<td>Summit Anesthesia Solutions, Ann Harbor, MI</td>
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			<td height="21" style="height:21px;">Specially designed animal holder</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
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			<td height="21" style="height:21px;">Bickford&#160;Omnicon F/Air&#160;anesthesia gas filter unit&#160;</td>
			<td>A.M. Bickford, Inc. Wales Center, NY</td>
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			<td height="21" style="height:21px;">Pulse-oximeter module, MR compatible small animal monitoring and gating system&#160;</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
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			<td height="21" style="height:21px;">Signal breaking module, MR compatible small animal monitoring and gating system</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
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			<td></td>
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			<td height="21" style="height:21px;">9.4 T (400 MHZ) 89 mm vertical core bore MR scanner&#160;</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
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			<td></td>
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			<td height="21" style="height:21px;">4-cm millipede micro-imaging RF coil&#160;</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
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			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">SAM PC monitor&#160;</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
			<td style="width:145px;"></td>
			<td></td>
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			<td height="21" style="height:21px;">Quantitative Medical Image analysis software&#160;</td>
			<td colspan="3">http://segment.heiberg.se;&#160; Segment v1.8 R1430,&#160; Medviso, Oresunds region, Sweden</td>
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			<td height="21" style="height:21px;">Matlab software&#160;</td>
			<td>The Mathworks, Inc.&#160; Natick, MA</td>
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			<td></td>
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			<td height="21" style="height:21px;">Computer-Unix operating system</td>
			<td style="width:364px;"></td>
			<td style="width:145px;"></td>
			<td></td>
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noninvasive assessment of cardiac abnormalities in experimental autoimmune myocarditis by magnetic resonance microscopy imaging in the mouse

心肌炎是心肌的炎症，但对疾病的影响的那些表演的临床表现只〜10％。研究心肌损伤的免疫事件，心肌炎的各种小鼠模型已被广泛使用。这项研究涉及实验性自身免疫性心肌炎（EAM）诱导的心肌肌球蛋白重链（MYHC）-α的A / J小鼠334-352段;受影响的动物培养淋巴细胞性心肌炎，但没有明显的临床症状。在这个模型中，磁共振显微镜（MRM）作为一种非侵入性方式的实用程序来确定显示在免疫的动物MYHC-α334-352心脏结构和功能改变。 EAM和健康的小鼠用9.4 T（400兆赫）89 mm垂直芯孔扫描仪配备了4厘米千足虫射频成像探头和100克/厘米三轴梯度成像。从麻醉动物用梯度回波为基础的电影脉冲序列被收购心脏图像，和阿尼玛LS是由呼吸和脉搏血氧饱和度监测。该分析显示，增加心室壁在EAM小鼠的厚度，并相应减少心室的内部直径，当与正常小鼠相比较。该数据表明，在发炎的心脏形态和功能变化可能是由MRM非侵入性地监测在活的动物。总之，MRM提供评估引起的感染性疾病的药物的心肌损伤的进展和回归，以及响应于治疗的优点。

在这份报告中，显示MRM技术的实用程序作为一种非侵入性的方式来确定受影响的与EAM动物的心的结构和功能改变。心肌炎诱导在A / J小鼠进行免疫的动物在CFA 7 MYHC-α334-352，并且将动物进行的MRM实验第21天免疫后。在活的动物异氟烷麻醉下使用89毫米垂直孔磁铁配备了三轴梯度（最大强度100克/厘米）的MRM成像进行9.4 T（400 MHz为质子）。球探图像被收购的定位和定位小鼠心脏中的视场的中心?...

Watch this Scientific Journal Video about Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse at JoVE.com

Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

心脏异常的实验性自身免疫性心肌炎的无创性评估磁共振成像显微镜在鼠标

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events ...

noninvasive-assessment-cardiac-abnormalities-experimental-autoimmune

Research

JoVE Journal

Medicine

9.8K 次观看.  University of Nebraska-Lincoln. This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

视频: Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

Magnetic Resonance Derived Myocardial Strain Assessment Using Feature Tracking

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF. Current technologies for CMR are time consuming and difficult to implement in clinical practice. Feature tracking is a technology that can lead to more automization and robustness of quantitative analysis of medical images with less time consumption than comparable methods.
Methods: An automatic or manual input in a single phase serves as an initialization from which the system starts to track the displacement of individual patterns representing anatomical structures over time. The specialty of this method is that the images do not need to be manipulated in any way beforehand like e.g. tagging of CMR images.
Results: The method is very well suited for tracking muscular tissue and with this allowing quantitative elaboration of myocardium and also blood flow.
Conclusions: This new method offers a robust and time saving procedure to quantify myocardial tissue and blood with displacement, velocity and deformation parameters on regular sequences of CMR imaging. It therefore can be implemented in clinical practice.

An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF.

磁共振衍生的心肌劳损评估使用特征跟踪

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that ...

科研

医学

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

利用磁共振成像，容积电脑断层扫描及超音波的裸鼠乳腺癌骨转移大鼠模型血管生成的多模态成像

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Diffusion Tensor Magnetic Resonance Imaging in the Analysis of Neurodegenerative Diseases

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present applications are designed to investigate differences of WM involvement patterns in different brain diseases, especially neurodegenerative disorders, by use of different DTI analyses in comparison with matched controls. 	
DTI data analysis is performed in a variate fashion, i.e. voxelwise comparison of regional diffusion direction-based metrics such as fractional anisotropy (FA), together with fiber tracking (FT) accompanied by tractwise fractional anisotropy statistics (TFAS) at the group level in order to identify differences in FA along WM structures, aiming at the definition of regional patterns of WM alterations at the group level. Transformation into a stereotaxic standard space is a prerequisite for group studies and requires thorough data processing to preserve directional inter-dependencies. The present applications show optimized technical approaches for this preservation of quantitative and directional information during spatial normalization in data analyses at the group level. On this basis, FT techniques can be applied to group averaged data in order to quantify metrics information as defined by FT. Additionally, application of DTI methods, i.e. differences in FA-maps after stereotaxic alignment, in a longitudinal analysis at an individual subject basis reveal information about the progression of neurological disorders. Further quality improvement of DTI based results can be obtained during preprocessing by application of a controlled elimination of gradient directions with high noise levels.	
In summary, DTI is used to define a distinct WM pathoanatomy of different brain diseases by the combination of whole brain-based and tract-based DTI analysis.

Diffusion tensor imaging (DTI) basically serves as an MRI-based tool to identify in vivo the microstructure of the brain and pathological processes due to neurological disorders within the cerebral white matter. DTI-based analyses allow for application to brain diseases both at the group level and in single subject data.

磁共振扩散张量成像在神经退行性疾病的分析

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present ...

Implanting Glass Spinal Cord Windows in Adult Mice with Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as multiple sclerosis. Here we present a low-cost and reproducible glass window implantation protocol that is suitable for intravital microscopy and studying the dynamics of spinal cord cytoarchitecture with subcellular resolution in live adult mice with EAE. Briefly, we surgically expose the vertebrae T12-L2 and construct a chamber around the exposed vertebrae using a combination of cyanoacrylate and dental cement. A laminectomy is performed from T13 to L1, and a thin layer of transparent silicone elastomer is applied to the dorsal surface of the exposed spinal cord. A modified glass cover slip is implanted over the exposed spinal cord taking care that the glass does not directly contact the spinal cord. To reduce the infiltration of inflammatory cells between the window and spinal cord, anti-inflammatory treatment is administered every 2 days (as recommended by ethics committee) for the first 10 days after implantation. EAE is induced only 2-3 weeks after the cessation of anti-inflammatory treatment.
Using this approach we successfully induced EAE in 87% of animals with implanted windows and, using Thy1-CFP-23 mice (blue axons in dorsal spinal cord), quantified axonal loss throughout EAE progression. Taken together, this protocol may be useful for studying the recruitment of various cell populations as well as their interaction dynamics, with subcellular resolution and for extended periods of time. This intravital imaging modality represents a valuable tool for developing therapeutic strategies to treat autoimmune demyelinating diseases such as EAE.

We describe a method for implanting and maintaining glass windows over the exposed spinal cords of adult mice for studying experimental autoimmune encephalomyelitis. These windows provide chronic optical access to the spinal cord for monitoring cell dynamics at the subcellular level in live animals using two-photon microscopy.

玻璃植入脊髓的Windows在成年小鼠实验性自身免疫性脑脊髓炎

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

同时PET / MRI成像在小鼠脑缺氧缺血

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

在横向主动脉缩窄的小鼠模型和心脏形态学评估功能改变通过超声心动图成像

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

通过轻型显微镜对小鼠急性心肌炎模型中白细胞浸润的定量可视化

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Cardiac Magnetic Resonance Imaging at 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher magnetic field strengths and potentially provides improved signal contrast and spatial resolution. While promising results have been achieved, ultra-high field CMR is challenging due to energy deposition constraints and physical phenomena such as transmission field non-uniformities and magnetic field inhomogeneities. In addition, the magneto-hydrodynamic effect renders the synchronization of the data acquisition with the cardiac motion difficult. The challenges are currently addressed by explorations into novel magnetic resonance technology. If all impediments can be overcome, ultra-high field CMR may generate new opportunities for functional CMR, myocardial tissue characterization, microstructure imaging or metabolic imaging. Recognizing this potential, we show that multi-channel radio frequency (RF) coil technology tailored for CMR at 7 Tesla together with higher order B0 shimming and a backup signal for cardiac triggering facilitates high fidelity functional CMR. With the proposed setup, cardiac chamber quantification can be accomplished in examination times similar to those achieved at lower field strengths. To share this experience and to support the dissemination of this expertise, this work describes our setup and protocol tailored for functional CMR at 7 Tesla.

The sensitivity gain inherent to ultrahigh field magnetic resonance holds promise for high spatial resolution imaging of the heart. Here, we describe a protocol customized for functional cardiovascular magnetic resonance (CMR) at 7 Tesla using an advanced multi-channel radio-frequency coil, magnetic field shimming and a triggering concept.

7号特斯拉的心脏磁共振成像

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

大鼠颈总动脉的相位对比磁共振成像

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Quantification of Levator Ani Hiatus Enlargement by Magnetic Resonance Imaging in Males and Females with Pelvic Organ Prolapse

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while evacuating acoustic gel, using a horizontally oriented 1.5 T magnetic resonance (MR) scanner. On midsagittal images, the vertical distance of pelvic organs is measured in millimeters relative to the hymen plane (female) and to the lower border of the symphysis pubis (male), preceded by - (above) or + ( below) signs. On axial images, the levator ani area is calculated in square centimeters with a free-hand tracing method from three key images, passing through the midsymphysis (level I), tangential to the lower border of the symphysis (level II), and at the maximal anterior rectal wall bulging (level III). Areas at rest and strained are compared to find evidence of a percentage of increase. The purpose is to provide objective evidence of the maximal extent of pelvic organs descent and hiatus enlargement without the interference of foreign objects or the examiner&#39;s proximity, so as to overcome the limitations of pelvic examination and transperineal sonography (i.e., subjectivity and sex-related limitations [female only]).

Here we present a protocol to standardize the measurement of the levator ani hiatus size&#160;by magnetic resonance imaging. The purpose is to extract biomechanical inferences from image analysis by comparing resting and straining values in patients of both sexes with pelvic prolapse, using consistent anatomical bony landmarks.

男性和女性盆腔器官脱垂的磁共振成像对左旋的定量研究

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while ...