这项工作得到了法国政府 (国家研究机构, 全国情报局) 的支持, 通过 "未来投资" LABEX SIGNALIFE 计划 (参考 ANR-11-LABX-0028-01) 和赠款的 k.d.w. 从协会倒 la 研究癌,法兰西基金会, 计划癌症 Inserm。库珀和影音从研究 Médicale 和尼斯城分别获得了奖学金。心动和换能器由飞利浦公司提供。我们感谢 a. Borderie、s Destree、m. Cutajar-Bossert、Landouar、Martres、Biancardini 和克里希纳瓦格纳的熟练技术援助。

此处所述的实验是按照相关的机构和法国动物福利法、指南和政策进行的。他们已获得法国伦理委员会 (Comit 和 #233; Institutionnel 和 #39; Ethique #39; 动物 de Laboratoire; 数字 NCE/2012-106). 
 1. 超声心动图 <ol> <li> 使用标准的实验室平衡来确定鼠标的体重, 同时将其轻轻地放在尾部以确保正确定位。
	<li> 麻醉通过腹腔 (ip) 注射50毫克/千克戊巴比妥 25 , 26 。
	注意: 如果在整个研究中使用相同的协议, 任何其他类型的麻醉都可以使用。下面将讨论优缺点. </li>
	<li> 将鼠标放回自己的笼子, 等到它没有反应, 它显示稳定的呼吸, 后足反射是缺席。为了测试这一点, 稍微挤压一只脚, 观察腿是否仍然缩回. </li>
	<li> 使用商用的啮齿动物剃须刀将胸腔和左腋窝的左侧剃须. 
	注意: 使用专用的啮齿动物剃须刀可以完全去除细小的小鼠毛, 以避免干涉超声心动图测量。商业脱毛膏或解决方案应避免, 因为它们通常是芳香, 这将扰乱动物后, 它唤醒。避免过度剃须, 因为它会增加热量损失. </li>
	<li> 将熟睡的动物放在一个暖垫上, 设置为 40-42 和 #176; C 在浅的左侧位置, 头部在 12 o 和 #39; 时钟和尾部在 6 o 和 #39; 时钟。用胶带固定左臂、左腿和尾部. </li>
	<li> 应用5ml 超声心动图凝胶到剃须的胸部和传感器的头部. </li>
	<li> 将传感器胸骨放在左侧, 将其定向到颈部的右侧, 以获得 two-dimensional (2D) 胸骨轴在乳突肌水平上的视面。转动传感器90和 #176; 顺时针方向以获得 papilary 肌肉水平的短视图。使用最小深度设置和缩放来最大化图像质量和帧速率。将扫描速度设置为最大值. 
	<ol> <li> 若要获取这些设置, 可以使用不同的缩放和深度设置选项, 具体取决于计算机和软件。在短视图 27 中记录 2 d 引导的 M 模式图像。请参阅 图 1A 和 B 27 . 
		注意: 小心避免对胸部施加过度压力, 因为这可能会导致心动过缓. </li>
	</ol> </li> <li> 记录每种动物至少3系列3心脏跳动的电影循环. 
	注: 对于本研究中使用的心动软件, 按 #34; 获取/保存/#34; 按钮一次。此方法特定于与此特定软件的心动。其他软件包可用于不同的计算机. </li>
	<li> 成功录音后, 从鼠标胸腔、发热垫和换能器中去除超声心动图凝胶。从四肢和尾部取下胶带. </li>
	<li> 将鼠标置于发热垫上, 用纸巾覆盖, 避免不必要的光照和热损耗, 直到它苏醒. </li>
	<li> 将动物放回笼子. </li>
	<li> 分析记录的 M 模式图像从胸骨短视图, 以确定左心室 (LV) 的尺寸和功能。在收缩期和舒张期 (LVAWs 和 LVAWd)、lv 内端收缩和舒张末期直径 (LVIDs 和 LVIDd) 和左心室后壁厚度 (LVPW) 的收缩和舒张期 (LVPWs 和 LVPWd) 中测量 lv 前壁的厚度。在存储的图像上标识组织-血液界面. </li>
	<li> 在 lv 后壁最前收缩期时, 测量表观最大左心室舒张维度和左心室终收缩维度时的舒张维度。点击触摸屏上的 #34; 分析和 #34; 图标, 然后在和 #34; LVAWd 和 #34; 图标。将电子卡尺定位于右心室腔与左前壁的舒张功能. </li>
	<li> 将电子卡尺放置在 lv 前壁和 lv 腔之间的接口上, 以获得 lv 舒张前壁厚度; 该软件将直接切换到 lv 内部舒张末期测量. </li>
	<li> 将卡钳放置在 lv 腔和 lv 后壁的接口上, 以获得 lv 内端舒张直径; 软件将切换到 lv 后壁厚度测量. </li>
	<li> 在 lv 后壁与心包的交界面上放置卡尺, 以获得左心室舒张后壁厚度。对于左心室收缩尺寸, 点击触摸屏上的和 #34; LVAWs 和 #34; 图标和位置在收缩期的右室腔和 lv 前壁的界面上的电子卡尺. </li>
	<li> 将电子卡尺放置在 lv 前壁和 lv 腔之间的接口上, 以获得 lv 收缩前壁厚度。重复上述过程, 为 lv 内端收缩直径和左心室收缩后壁厚度。使用美国超声心动图学会采用的前沿公约跟踪心内膜和心外膜边界 13 , 27 。
	注: LV 收缩功能参数将使用以前的测量自动计算。LV 分数缩短 (fs) 定义为 fs (%) = [(LVIDd-LVIDs)/LVIDd] x 100。LV 弹出分数 (ef) 是用修改后的 Teicholz 公式计算的, 其中 ef (%) = [(LVIDd 3 -LVIDs 3 )/LVIDd 3 ) x 100 12 。请参阅 图 1A 和 B 。
	<li> 将数据存储在光盘或 USB 内存棒上, 并制作备份副本. </li>
	<li> 使用适当的软件导入、分析和导出数据. 
	注意: 在这些基线测量之后, 可以暂停实验。如果对击倒动物和野生型窝进行直接比较, 请进行组织学分析 6 。Cre-ERT2;液氧/液氧鼠标线, 继续第二天与他莫昔芬诱导的 ip 注入, 如描述的 5 , 24 。如果通过结扎左冠状动脉来诱发实验性心肌梗死 5 , 28 , 手术可以直接在超声心动图测量后进行, 当小鼠都在麻醉之下否则, 应维持两轮麻醉之间的最短延迟一周, 以限制术后致死率. </li>
</ol> 2。组织学评价用心脏标本的制备 <ol> <li> 用颈椎脱位处死动物。测量他们的身体重量。用70% 的酒精拭子消毒 #160; 胸部和腹部. </li>
	<li> 在胸骨远端1厘米的皮肤上做一个横切口。使用钝钳, 从胸腔中取出皮肤, 向头部方向移动。用细钳轻轻握住胸骨, 用细剪刀的钝端打开隔膜. </li>
	<li> 将两侧的肋骨固定在胸骨上。向头部方向移动胸骨。在胸腔找到心脏用细钳夹住心脏的血管主干, 然后用细剪刀切割. </li>
	<li> 打开胸腔并切除整个心脏, 测量心脏重量, 并建立一个 heart-to-body 重量比 4 , 5 , 6 , 23 , 29 , 30/我的 </li>
	<li> 将2毫升10% 中性缓冲福尔马林溶液中的心脏固定在15毫升的管子中过夜, 在4和 #176; C. 
	小心: 危险!使用福尔马林溶液的工作必须在化学油烟罩中进行;戴上手套和安全眼镜. 
	注: 由于福尔马林溶液的缓冲成分因不同的供应商而异, 因此在整个研究中使用相同的供应商. </li>
	<li> 第二天, 在横切面上切开心脏, 在中间, 然后将它们转换成石蜡包埋盒, 这是在病理实验室使用自动嵌入装置进行的. </li>
	<li> 执行切片. 
	<ol> <li> 节石蜡块的厚度为3和 #181; m 使用切片, 并将其浮在40和 #176 中; C 水浴含蒸馏水. </li>
		<li> 将稿件转移到幻灯片上。允许幻灯片在37和 #176 中过夜; c 孵化器, 并将它们存储在4和 #176; c 直到准备就绪. 
		注意: 可以在此处暂停协议, 直到用户准备好染色 (步骤 3). </li>
	</ol> </li> <li> Deparaffinize 和水化组织幻灯片. 
	<ol> <li> 将幻灯片放入着色罐中, 并在55和 #176 中放置玻璃插入物; C 烤箱10分钟融化石蜡. </li>
		<li> Deparaffinize 将200毫升二甲苯或二甲苯的两个变化中的幻灯片替换为5分钟. 
		警告: 高度易燃和有毒!在化学油烟罩中工作;戴上手套和安全眼镜. </li>
		<li> 将幻灯片传送到200毫升的100% 酒精。做两个变化3分钟, 并转移一次通过200毫升95% 酒精3分钟. 
		警告: 高度易燃!远离点火源;禁止吸烟. </li>
		<li> 在200毫升磷酸盐缓冲盐水溶液 (PBS) 中冲洗两次, 每次5分钟, 然后继续步骤3、4或 5. </li>
	</ol> </li> </ol> 3。苏木精和曙红染色 <ol> <li> 在蒸馏水中用它们的剖面冲洗幻灯片. </li>
	<li> 用苏木精溶液将细胞核染色8分钟 </li>
	<li> 在自来水中冲洗10分钟. </li>
	<li> 用曙红溶液染色2分钟 </li>
	<li> 脱水三次, 2 分钟, 100% 乙醇 (乙醇)。二甲苯或二甲苯代用品2分钟清除三次。安装在 xylene-based 安装介质中. </li>
	<li> 照片的幻灯片和测量的心肌细胞直径在室隔膜的纵向部分的核心水平。每节至少测量100细胞, 每心脏三节. </li>
</ol> 4。WGA 染色 <ol> <li> 用甲基 (或其他荧光染料) 孵育从步骤2.7 获得的幻灯片--共轭 WGA (PBS 中的 1:100), 在潮湿的室内温度为60分钟. </li>
	<li> 用 PBS 洗涤三次, 每次5分钟. </li>
	<li> 装有带有 DAPI 的荧光安装介质的安装。存储在4和 #176; C 在黑暗中在分析之前. 
	注意: 佩戴防护眼罩和兼容的耐化学手套. </li>
	<li> 拍摄幻灯片. 
	<ol> <li> 使用带有荧光 epi 照明的显微镜和 DAPI 和甲基的滤镜集 (请参阅 材料表 )。将光圈设置为最大值和亮度为曝光. </li>
		<li> 获取蓝色和红色通道的400x 放大倍数的单独图像。打开 ImageJ 中的图像。根据需要调整亮度和对比度 (图像和 #62; 调整和 #62; 亮度/对比度). </li>
		<li> 将每个图像设置为8位 (图像和 #62; 类型和 #62; 8 位)。覆盖 DAPI 和 WGA 图像。使用蓝色通道为 DAPI 和红色渠道为 WGA;将绿色和灰色通道设置为 none (图像和 #62; 颜色和 #62; 合并通道) 31 . </li>
	</ol> </li> <li> 确定室隔膜横截面上的细胞核水平的心肌细胞直径。
	<ol> <li> 定义 ImageJ 中图像的比例。为此, 拍摄已知大小的对象 ( 例如, 与心脏部分相同的放大倍数的例室; 步骤 4.4). </li>
		<li> 使用 straight-line 工具从已知结构的开始到结束处绘制直线 (分析和 #62; 设置刻度). 
		注意: 以像素为单位的距离将自动显示;必须输入已知的距离和长度单位。在细胞核的水平上进行心肌细胞的测量. </li>
		<li> 通过 DAPI 阳性核向 WGA 阳性细胞膜 (分析和 #62; 测量) 的相反位置绘制一条从 WGA 阳性膜的直线。在 "结果" 窗口中, 确保长度值有一个有意义的小数位 (分析和 #62; 设置度量和 #62; 小数位置). </li>
		<li> 每节至少测量100单元格, 每个心脏三节。将结果导出到 Excel (单击结果和 #62; 文件和 #62; 保存为和 #62;. csv) 6 , 31 . </li>
	</ol> </li> </ol> 5。Pecam-1 免疫 <ol> <li> 执行抗原揭露. 
	<ol> <li> 将1600毫升柠檬酸钠缓冲液 (10 毫米柠檬酸钠, 0.05% 吐温 20, pH 6.0) 添加到压力锅中。将压力锅放在板上, 并将其完全通电。在这一点上, 不要保证压力锅的盖子;简单地在上面休息. 
		注意: 热! </li>
		<li> 一旦柠檬酸钠缓冲液沸腾, 将幻灯片从步骤2.5 转移到压力锅。保护压力锅盖。一旦炊具已达到全部压力, 等待7分钟。当7分钟已经过去了, 关闭板和放置高压锅在一个空水槽。启动压力释放阀, 在炊具上运行冷水。一旦它有 de-pressurized, 打开盖子和运行冷水到炊具5分钟, 放置在200毫升 PBS 的幻灯片. 
		注: 另外, 可使用微波抗原揭露, 虽然过热的风险增加. </li>
	</ol> </li> <li> 在甲醇中使用0.3% 过氧化氢来阻止内源性过氧化物酶活性为5分钟. 在200毫升的 PBS 中冲洗三次, 每张2分钟. 
	警告: 易燃和有毒! </li>
	<li> 在稀释的正常阻断血清 (PBS 中5% 正常山羊血清) 中孵育15分钟的幻灯片, 其中也含有素块 (4 滴1毫升). </li>
	<li> 小心地从切片中挖掘液体, 并用家兔的 Pecam-1 抗体孵育它们, 在 PBS 中稀释 1:50, 其中含有2.5% 正常山羊血清和4滴生物素块每毫升。在4和 #176 的潮湿的室内, 在一夜之间孵化幻灯片; C. </li>
	<li> 在 PBS 200 毫升中每5分钟清洗一次幻灯片三次。在室温下, 用化山羊兔 IgG 抗体稀释1:200 的 PBS 中含有2.5% 正常山羊血清1小时。在200毫升 PBS 中每三次洗涤5分钟的幻灯片. </li>
	<li> 孵化 t他的部分与素/biotin-based 过氧化物酶系统20分钟 (试剂 A 和试剂 B 需要结合30分钟之前使用)。在200毫升 PBS 中每三次洗涤5分钟的幻灯片. </li>
	<li> 溶解 1 33 和 #39;-diaminobenzidine (民建联) 和1尿素过氧化氢片剂5毫升的双水。用民建联的溶液孵育约3分钟, 仔细监测颜色的发展。通过在200毫升 PBS 中轻轻地冲洗幻灯片来停止颜色反应. 
	警告: 致癌;穿防化学手套! </li>
	<li> 用苏木精将细胞核 Counterstain 6 分钟。在自来水中冲洗2分钟脱水三次, 2 分钟, 每200毫升100% 酒精。在200毫升的二甲苯或二甲苯替代品中, 每2分钟清除三次。安装在 xylene-based 安装介质中. </li>
	<li> 在每颗心脏的室间隔40x 倍的情况下拍摄幻灯片 (至少十字段), 并使用自由可用的 ImageJ 软件 31 来测量 Pecam-1 区域的密度。使用颜色反褶积 32 插件, 为民建联和苏木精和调整图像对比度相同的水平. 
	注意: 如果褐色和紫色/蓝色颜色有明显的光谱重叠, 可能会导致颜色反褶积困难, 你可以尝试不同品牌的苏木精溶液, 以获得明确的, 蓝色核染色。或者, 可以对毛细血管进行免疫荧光或人工计数. </li>
</ol>

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作者没有什么可透露的。

对小鼠心脏结构和功能进行了不同的评价, 包括超声心动图、造影增强 MRI、微 CT 和 PET 扫描。由于其 cost-effectiveness 和简单, 超声心动图是最广泛使用的技术, 功能分析小鼠11。一般来说, 由于心脏的小尺寸和心率的高频率在小鼠, 传感器以频率和 #62; 10 兆赫应该使用, 虽然成功的测量被报告了与8或9兆赫传感器4,7.由于心功能与体温和心脏频率密切相关, 因此在整个研究过程中控制这些参数是很重要的。将鼠标放在加热垫上是保持麻醉动物体温恒定的必要条件。理想情况下, 应使用一个带有直肠探头的可控加热垫, 将体温设置为38° c。如果这是不可用的, 暖气垫应设置为二至三度以上的这个值。应避免加热灯, 因为它们使调查员的任务复杂化, 并有可能使动物过热。
为了控制心率和心功能, 麻醉的类型是非常重要的。大多数研究使用异氟醚, 因为麻醉是容易诱导吸入在诱导室 (3% 异氟醚), 可以通过吸入面罩 (1-2% 异氟醚), 并仅是短时间内维持。其他常用物质有 22, 2-tribromoethanol, 戊巴比妥和氯胺酮 + 嗪混合11,12。令人惊讶的是, 据报道, 异氟醚在超声心动图测量中损害心脏功能, 这种作用在氯胺酮 + 嗪组11中更为明显。单氯胺酮在这项研究中效果最好11。高et al。报告了具有22、2-tribromoethanol12的最可重现的结果。结果在相同的范围内的异氟醚, 22, 2-tribromoethanol, 戊巴比妥12。此外, 22、2-tribromoethanol 和戊巴比妥组的心率也不同,12。心脏et al.报告氯胺酮 + 嗪组的心率降低, 与22、2-tribromoethanol 组16相比较。在我们的实验中使用不同的转基因小鼠和戊巴比妥麻醉, 平均心率范围介于350和 450 bpm 之间。然而, 弹射分数是和 #62; 80%, 对应于价值在神志清楚的动物11。超声心动图测量在基线条件下从我们的实验室4,56与其他地方报告的范围相同111214 ,15,16,171819202122.与戊巴比妥, 氯胺酮, 异氟醚和 22, 2-tribromoethanol 的特点是快速发作和恢复。因此, 在麻醉和超声心动图测量之间的时间应该是相同的所有动物的研究, 以避免不同程度的恢复从麻醉。戊巴比妥的作用更长, 但有一个小的治疗窗口的缺点, 使它有必要调整剂量紧与体重有关。长期麻醉使用戊巴比妥的优势, 在基线超声心动图测量, 手术程序可以执行, 而无需回5。应避免使用戊巴比妥的重复麻醉, 因为根据我们的经验, 注射时间间隔少于一周导致术后死亡率增高。
在与 #62 的小鼠进行超声心动图时; 10 兆赫传感器, 图片的质量应足以确定全局 LV 功能。为了减少心跳变异和运动伪影, 必须为每种动物拍摄几张照片。专家的意见, 从训练有素的心脏病学家或科学家的经验, 在小鼠的超声心动图应该在开始时, 以评估质量的图像。大多数超声心动图机使用 Teicholz 公式11,12从 LV 内径计算 EF。即使在 lv 尺寸很容易测量, 他们提供了一个不完善的评估 lv 体积和区域, 因为 lv 不符合任何理想, 简化几何形状。Teicholz 公式获得的分数缩短和射出分数都是确定 LV 收缩函数的完美参数, 因为它们都是基于几何假设, 并且只描述了两个壁的收缩力。用双辛普森的方法评估的射血分数更准确, 但这是不可能得到的每一个动物用这里使用的设备。
心肌梗死后, 我们面临着与超声心动图成像有关的几个问题。首先, 开胸疤痕严重阻碍了图像质量的提高。其次, 动物对重复麻醉更敏感。最后, M-c/s lv 容量和作用测量假设 lv 几何是同类的。因此, 在心肌梗死后的 LV 壁运动异常的存在下, 这些指标的使用变得更加有争议。
在我们的手中, 使用一个15兆赫探头允许偶尔, 但不是系统的多普勒图像记录。需要更高的频率, 以确定 LV 区域功能或执行心肌声学造影, 只能获得与专用于啮齿目动物的设备12。
考虑到这些啮齿动物特有的系统有更多的可能性和更高的分辨率, 我们应该在将来使用它们。缺点是更高的费用和一个专用空间的需要。这些高端, 啮齿动物专用的 echocardiographs 将是有利可图的, 只有当有足够的动物正在调查和关键数量的科学家使用的设备。通过为这些机器提供的特殊训练, 他们可以被科学家们用来做心脏病学专家。临床超声心动图设备提供了一个有限的解决方案, 如上所述。然而, 它仍然成功地用于不同的经验 laboratories11,12;可以很容易地被熟练的科学家和心脏病专家使用;对专用空间的移动和要求更低;而且, 由于机器数量很大, 允许考虑不同的选择来降低成本 (例如,租用, 获得不在临床使用的设备, 或进行二手采购)。
对于组织学分析, 第一个关键点是减少器官隔离和组织固定之间的时间。正如本议定书, 大多数染色和组织学分析是基于透射光显微镜;甲醛心脏组织的浸入式固定是足够的。如果一个项目的重点是更多的荧光显微镜染色, 应该考虑灌注固定的麻醉动物, 以消除红细胞的组织, 这表明明亮的荧光。
为了避免石蜡嵌入的变化, 我们使用自动嵌入装置。由于石蜡的熔点和硬度不同, 我们建议在整个研究中使用相同的供应商。石蜡切片应进行 pre-chilled 块使用旋转切片。截面厚度必须保持不变。3-µm 剖面厚度允许清晰可视化的膜边界在苏木精-曙红染色心脏切片, 这是需要准确测量心肌细胞直径。由于心肌细胞的形状不规则, 必须在细胞核的水平上进行测量。WGA 染色提供了一种替代方法, 染色的细胞膜, 将导致与苏木素-曙红染色相同的价值。在形态分析之前, 你应该清楚地定义心脏的哪个部分 (即,左或右心室壁或隔膜) 将被测量并且心肌细胞横断面是否在长或短的轴确定。由于心肌细胞的横向、螺旋和纵向方向, 使心肌细胞在同一横断面上获得了可能。是否测量纵向或横向直径是一个惯例问题, 只要在细胞核的水平上分析细胞。
在我们的研究中, 我们观察到的超声心动图尺寸和 heart-to-body 体重比率和心肌细胞大小之间的协议4,5,6, 但心肌细胞大小的组织学测量不始终对应于射出分数的值。例如, 运动训练会增加心脏尺寸, 保留射血分数和增加心肌细胞直径。然而, 失代偿性心力衰竭的特点是增加心脏尺寸, 降低射血分数和增加心肌细胞直径33。此外, 我们最近发现, 由于血管内皮特异性过度表达而增加的心脏血管密度不会在基线条件下改善心脏功能, 也不会导致心肌梗死后的 PPARβ恢复,5.
对于免疫组化, 重要的是要包括负控制和执行在协议中提供的不同的阻断步骤。该协议的抗原揭露步骤是特定的主要抗体使用。对于不同的主抗体, 有必要确定或 high-pH 揭露缓冲器是否能产生更好的信号。不同的固定技术可用于检测特定抗原。例如, 据报道, Pecam-1 标记在锌固定、石蜡包埋的组织中最有效, 而同样的固定需要额外的步骤来减少血栓调节蛋白抗体34的背景。因此, 我们使用定期福尔马林固定, 使研究扩展到各种不同的抗原。替代 Pecam-1 免疫, isolectin B4 经常用于可视化容器。除了内皮细胞的比例35外, isolectin B4 标签还胶质胶36和巨噬细胞37。此外, 各种抗原可用于区分动脉和静脉内皮细胞38。一个优雅的方式来可视化功能灌注血管或确定血管发芽或回归是静脉注射荧光耦合凝集素, 然后分析 cryosections24。然而, 这种方法在很大程度上限制了检测额外抗原的可能性, 并由于 cryosections 的质量不同而进行形态分析。
在 immunostained 部分的信号是用民建联的可视化, 面积密度可以定量确定使用自由可用的 ImageJ 软件。然而, 由于信号不是线性的, 褐色染色的程度不完全对应于蛋白质表达水平。

有大量的转基因小鼠模型可供使用, 而在老鼠体内药理学研究的数量是高1,2。超声心动图和 MRI 是常用的方法的表型表征心脏功能和形态学在这些小鼠模型3。该方案的目的是分析成年小鼠的心功能和形态学。它结合了超声心动图、组织学和免疫组化测量。超声心动图检测在小鼠中广泛应用4,56,78910 12。Pachon et al.11已确定在循环中发布的205项研究,循环研究,美国生理学杂志-心脏和循环系统生理学, 和心血管研究之间的2012和2015用超声心动图检查动物。
超声心动图用于鉴别基因修饰小鼠心脏表型5,6,13,1415,1617,18,19,20,21,22, 以及分析小鼠慢性超负荷诱发肥大、心肌缺血和心肌病模型的心功能 (在12中进行了回顾)。改进的超声心动图设备允许对左心室收缩和舒张维度、组织多普勒成像、心肌对比声像的标准测量, 以及对 lv 区域功能和冠脉储备的评价12. 理想情况下, 应在有意识的小鼠中进行超声心动图检查, 以避免麻醉对收缩功能、自主反射控制和心率的负面影响11。然而, 这种方法受训练动物的要求的限制;保持体温稳定的困难;运动工件;应力;非常高的心脏频率;并且要求至少两名调查员进行实验, 特别是在大量动物被调查的情况下。有趣的是, 最近的一项研究报告在经过训练和未经训练的动物中, 超声心动图参数没有差异19。我们在麻醉小鼠身上进行超声心动图测量。下面将讨论不同的麻醉方案。
虽然标准分辨率超声心动图 (和 #62; 10 兆赫) 是足够的测量左心室收缩和舒张功能的成年小鼠, 该方法是有限的, 其基本结构现象的描述。因此, 我们结合的在体内测量与组织学和免疫的分析, 以测量, 例如, 心肌细胞直径和血管密度。其他组织学和免疫的研究, 如测定增殖, 细胞凋亡的检测, 梗死面积的测量, 纤维化的测定, 和特定的标记表达, 也可以进行相同类型的加工过的组织, 但不是本议定书的主题。体内超声心动图与组织学分析相结合, 为基础结构改变提供了更多的见解。另外一步, 我们可以通过分子和微的调查来完成这些测量。组织学分析不仅完成了超声心动图检查, 而且在超声心动描记术的分辨率不足时也成为必不可少的。这是特别的情况下, 在模型的转基因小鼠是胚胎致命的23,24。

<table>
	<tbody>
		<tr>
			<td>Wheat germ agglutinin (WGA) conjugated tetramethylrhodamine</td>
			<td>Life Technologies, Molecular Probes</td>
			<td>W849</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotinylated Goat Anti-Rabbit IgG Antibody</td>
			<td>Vectorlabs</td>
			<td>BA-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Avidin/Biotin Blocking Kit</td>
			<td>Vectorlabs</td>
			<td>SP-2001</td>
			<td></td>
		</tr>
		<tr>
			<td>VECTASTAIN Elite ABC HRP Kit (Peroxidase, Standard)</td>
			<td>Vectorlabs</td>
			<td>PK-6100</td>
			<td></td>
		</tr>
		<tr>
			<td>VECTASHIELD Antifade Mounting Medium with DAPI</td>
			<td>Vectorlabs</td>
			<td>H-1200</td>
			<td></td>
		</tr>
		<tr>
			<td>SIGMAFAST 3,3&#39;-Diaminobenzidine tablets</td>
			<td>Sigma</td>
			<td>D4168</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrogen peroxide solution</td>
			<td>Sigma</td>
			<td>H1009</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Pecam-1 (CD31) antibody</td>
			<td>Abcam</td>
			<td>ab28364</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound transmission gel, Gel Aquasonic 100</td>
			<td>Parker</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Linear ultrasound probe, L15-7io</td>
			<td>Philips Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Echocardiograph, IE33 xMATRIX</td>
			<td>Philips Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope, Leica DMi8</td>
			<td>Leica</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence Filterset DAPI</td>
			<td>Leica</td>
			<td>11525304</td>
			<td></td>
		</tr>
		<tr>
			<td>Filterset TxR</td>
			<td>Leica</td>
			<td>11525310</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital Camera, SPOT RT3 Color Slider</td>
			<td>Spot Imaging</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging Software, SPOT 5.2 Advanced and Basic Software</td>
			<td>Spot Imaging</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging Computer</td>
			<td>Dell</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Scissors</td>
			<td>Fine Science Tools</td>
			<td>14028-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Large Scissors</td>
			<td>Fine Science Tools</td>
			<td>14501-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel blades</td>
			<td>Fine Science Tools</td>
			<td>10023-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>Fine Science Tools</td>
			<td>11650-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Rodent shaver</td>
			<td>Harvard Apparatus</td>
			<td>34-0243</td>
			<td></td>
		</tr>
		<tr>
			<td>cassettes for paraffin embedding</td>
			<td>Sakura</td>
			<td>4155F</td>
			<td></td>
		</tr>
		<tr>
			<td>neutral buffered Formalin</td>
			<td>Sakura</td>
			<td>8727</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Sakura</td>
			<td>8733</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraffine TEK III</td>
			<td>Sakura</td>
			<td>4511</td>
			<td></td>
		</tr>
		<tr>
			<td>automated embedding apparatus, Tissue-Tek VIP</td>
			<td>Sakura</td>
			<td>6032</td>
			<td></td>
		</tr>
		<tr>
			<td>paraffin-embedding station Tissue-Tek TEC 5</td>
			<td>Sakura</td>
			<td>5229</td>
			<td></td>
		</tr>
		<tr>
			<td>microtome blades,Accu-Edge S35</td>
			<td>Sakura</td>
			<td>4685</td>
			<td></td>
		</tr>
		<tr>
			<td>microscopy slides, Tissue-Tek</td>
			<td>Sakura</td>
			<td>9533</td>
			<td></td>
		</tr>
		<tr>
			<td>cover slips, Tissue-Tek</td>
			<td>Sakura</td>
			<td>9582</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounting medium Tissue-Tek</td>
			<td>Sakura</td>
			<td>1408</td>
			<td></td>
		</tr>
		<tr>
			<td>slide boxes</td>
			<td>Sakura</td>
			<td>3958</td>
			<td></td>
		</tr>
		<tr>
			<td>eosine solution</td>
			<td>Sakura</td>
			<td>8703</td>
			<td></td>
		</tr>
		<tr>
			<td>hematoxyline solution</td>
			<td>Sakura</td>
			<td>8711</td>
			<td></td>
		</tr>
		<tr>
			<td>microtome, RM2125RT</td>
			<td>Leica</td>
			<td>720-1880 (VWR)</td>
			<td></td>
		</tr>
		<tr>
			<td>water bath, Leica HI1210</td>
			<td>Leica</td>
			<td>720-0113(VWR)</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>VWR</td>
			<td>ACRO444220050</td>
			<td></td>
		</tr>
		<tr>
			<td>15 ml tubes</td>
			<td>VWR</td>
			<td>734-0451</td>
			<td></td>
		</tr>
		<tr>
			<td>staining glass dish</td>
			<td>VWR</td>
			<td>MARI4220004</td>
			<td></td>
		</tr>
		<tr>
			<td>staining jars</td>
			<td>VWR</td>
			<td>MARI4200005</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>Binder</td>
			<td>9010-0012</td>
			<td></td>
		</tr>
		<tr>
			<td>DAB and urea hydrogen peroxide tablets, SIGMAFAST 3,3&#8242;-Diaminobenzidine tablets</td>
			<td>Sigma</td>
			<td>D4293</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS (10X)</td>
			<td>Thermo Fisher Scientific</td>
			<td>70011044</td>
			<td></td>
		</tr>
	</tbody>
</table>

echocardiographic and histological examination of cardiac morphology in the mouse

近年来, 越来越多的基因改良的小鼠模型已成为可用。此外, 在小鼠体内进行药理学研究的数量也很高。这些小鼠模型的表型特征也需要检查心脏功能和形态学。超声心动图和磁共振成像 (MRI) 是常用的方法来表征心脏功能和形态学的小鼠。专门用于小型啮齿动物的超声心动图和 MRI 设备昂贵, 需要专用空间。该协议描述了使用15兆赫人体血管探针的临床超声心动图系统对小鼠的心脏测量。对麻醉成年小鼠进行了测量。在胸骨短视图中, 每种动物在 M 模式下记录和分析至少三图像序列。术后进行心脏组织学检查, 在苏木素-曙红-或麦胚凝集素 (WGA) 染色石蜡切片上测定心肌细胞直径。Pecam-1 免疫后, morphometrically 测定血管密度。该协议已成功应用于基线条件下的药理学研究和不同的遗传动物模型, 以及经永久性结扎左前降冠状动脉后的实验性心肌梗死后 (小伙子)。根据我们的经验, 超声心动图的研究仅限于麻醉动物, 在体重至少25克的成年小鼠中是可行的。

在图 1中, 有代表性的超声心动图显示了超声心动描记对转基因小鼠心脏表型的鉴别作用。有正常心功能的小鼠 (图 1A) 和左心室扩张的动物和减少的 LV 功能 (图 1B) 之间的区别可以很容易地被识别出来。图 2显示了在没有 (图 2A) 的动物中进行心肌细胞直径测量的比较 (图 2B), 使用苏木精-曙红染色石蜡切片和测量心脏组织的纵断面。图 3演示了使用控制小鼠 (图 3A) 和心脏肥大的动物 (图 3B) 的心 WGA 染色石蜡切片测量心肌细胞横向直径的方法。图 4描述了心脏切片的 Pecam-1 免疫组织化学 (民建联、褐色染色) 的效用, 以确定正常心脏的血管密度 (图 4A) 和增加血管生成的心脏 (图4B).
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55843/55843fig1.jpg"> 
图 1: 标准分辨率超声心动图 (和 #62; 10 兆赫), 测量心肌梗死后扩张型心肌肥厚小鼠的左心室收缩和舒张维度及心功能与对照动物。 典型的超声心动图记录正常, 健康小鼠 (A) 和动物的 lv 扩张和减少左心室功能后心肌梗塞 (Tie2-CreERT2;PPARβ/δ + 他莫昔芬鼠)5 (B)。LVAW, LVID 和 LVPW 被表明。白条代表收缩和舒张的测量点。它们对应于为度量设置的游标点, 并在协议步骤1.12 中解释。<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55843/55843fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55843/55843fig2.jpg"> 
图 2: 苏木精-曙红-红染色切片, 用于测量扩张型心肌肥厚和控制动物的小鼠纵向切片细胞的心肌肌直径.正常心肌细胞大小 (A) 和心肌细胞大小增加的小鼠的心肌细胞直径的代表性图像 (Tie2-CreERT2;PPARβ/δ + 他莫昔芬鼠)5 (B)。黑线表示测量的直径。每个测量的值都表示。缩放栏 = 50 µm.<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55843/55843fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55843/55843fig3.jpg"> 
图 3: WGA 染色切片, 用于测量心肌肥厚小鼠与对照动物心脏组织部分的肌细胞横向直径.正常心肌细胞大小 (A) 和心肌细胞大小增加的小鼠的心肌细胞直径的代表性图像 (Tie2-CreERT2;PPARβ/δ + 他莫昔芬鼠)5 (B)。细胞核与 DAPI 1-4。白线表示在原子核 (蓝色) 的水平上测量的直径。每个测量的值都表示。缩放栏 = 50 µm. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55843/55843fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55843/55843fig4.jpg"> 
图 4: Pecam-1 immunolabeled 切片, 以分析控制小鼠和动物血管增生的心脏血管密度。 具有正常血管密度 (A) 和增强血管的动物 (Tie2-CreERT2) 血管标记的典型图像PPARβ/δ + 他莫昔芬鼠)5 (B)。细胞核与苏木精1-4。箭头指向 Pecam-1-positive 的容器。缩放栏 = 50 µm.<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55843/55843fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse at JoVE.com

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

超声心动图检查常用于小鼠。为此, 开发了昂贵的高分辨率超声波装置。该协议描述了一个负担得起的超声心动图方法结合组织学形态学分析, 以确定心脏形态学。

超声心动图与小鼠心脏形态学的组织学检查

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

echocardiographic-histological-examination-cardiac-morphology

Nanyang Technological University

Research

JoVE Journal

Medicine

21.2K 次观看.  CNRS, INSERM, iBV. 超声心动图检查常用于小鼠。为此, 开发了昂贵的高分辨率超声波装置。该协议描述了一个负担得起的超声心动图方法结合组织学形态学分析, 以确定心脏形态学。

视频: Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

使用小动物看更衣室反转恢复（SALLI）的MRI大鼠心脏功能和心肌形态学评估

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

科研

医学

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

右心小鼠的超声心动图评价

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled microscopic lesions (microlesions) that are remarkable due to the absence of infiltrating immune cells. Due to their release of cardiotoxic products, S. pneumoniae within microlesions are thought to contribute to the heart failure that is frequently observed during fulminate invasive pneumococcal disease in adults. Herein is demonstrated a protocol for experimental mouse infection that leads to reproducible cardiac microlesion formation within 30 hr. Instruction is provided on microlesion identification in hematoxylin &#38; eosin stained heart sections and the morphological distinctions between early and late microlesions are highlighted. Instruction is provided on a protocol for verification of S. pneumoniae within microlesions using antibodies against pneumococcal capsular polysaccharide and immunofluorescent microscopy. Last, a protocol for antibiotic intervention that rescues infected mice and for the detection and assessment of scar formation in the hearts of convalescent mice is provided. Together, these protocols will facilitate the investigation of the molecular mechanisms underlying pneumococcal cardiac invasion, cardiomyocyte death, cardiac remodeling as a result of exposure to S. pneumoniae, and the immune response to the pneumococci in the heart.

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

可视化<em&gt;肺炎链球菌</em&gt;在心脏Microlesions和随后的心脏重构

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

Do's and Don'ts in the Preparation of Muscle Cryosections for Histological Analysis

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of neuromuscular disorders. However, in order to do so, proper care needs to be taken when excising and preserving tissues to achieve optimal staining. One method of tissue preservation involves fixing tissues in formaldehyde and then embedding them with paraffin wax. This method preserves morphology well and allows for long-term storage at RT but is cumbersome and requires handling of toxic chemicals. Further, formaldehyde fixation results in antigen cross-linking, which necessitates antigen retrieval protocols for effective immunostaining. On the contrary, frozen sectioning does not require fixation and thus retains biological antigen conformation. This method also provides a distinct advantage in quick turn around time, making it especially useful in situations needing quick histological evaluation like intraoperative surgical biopsies. Here we describe the most effective method of preparing muscle biopsies for visualization with different histological and immunological stains.

Here we demonstrate the most efficient methods for freezing, embedding, cryosectioning, and staining of muscle biopsies to avoid freezing artifacts.

该做什么和不该做什么的肌肉冰冻切片的准备组织学分析

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of ...

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 ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

解剖的小鼠胰腺的组织学分析与代谢谱

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

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 ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV diastolic function is rarely quantified in experimental studies. This might be due to technical difficulties in the visualization of the RV in the apical four-chamber view in rodents. Here we describe two positions facilitating the visualization of the apical four-chamber view in mice to assess the RV diastolic function.
The apical four-chamber view is enabled by tilting the mouse fixation platform to the left and caudally (LeCa) or to the right and cranially (RiCr). Both positions provide images of comparable quality. The results of the RV diastolic function obtained from two positions are not significantly different. Both positions are comparably easy to perform. This protocol can be incorporated into published protocols and enables detailed investigations of the RV function.

Here we describe and compare two positions for obtaining the apical four-chamber view in mice. These positions enable the quantification of the right ventricular function, provide comparable results, and can be used interchangeably.

小鼠右心室舒张参数的超声心电图测量

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

兔子模型中的跨胸超声心动图检查

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...