我们要感谢科里博士雷诺兹和他们的专家的帮助，这个项目和实物设施的表型核心BCM的鼠标。 XHTW是WM凯克基金会杰出青年学者在医学研究中，也是由NHLBI /美国国立卫生研究院拨款HL089598 R01 - R01 - HL091947，和肌肉萎缩症协会授予＃69238支持。这项工作还支持在CaMKII信号在心脏基金会Leducq联盟的一部分。

1。影像学的准备小鼠<ol><li>之前，影像学，麻醉诱导室的鼠标（2％异氟醚0.5 L / min的混合100％O 2 ）。确保废气过滤器的操作人员的安全。从诱导室取出的动物，并用推剪剃的皮毛，从领口到中期胸部水平。然后取出剩余的体毛用脱毛膏。这种准备可能前一天进行影像学检查，以尽量减少在小鼠体内潜在的不受欢迎的应激反应。 </li><li>应用duralube凝胶双眼，以防止巩膜干燥（凝胶撒施，避免接触角膜）。麻醉鼠标放置在一个与嵌入式心电加热垫之上仰卧位信息，以保持体温。 </li><li>将鼻锥内麻醉系统连接到整个过程中保持稳定状态的镇静水平（1.0％至1.5％异氟醚0.5 L / min 的 100％O 2的混合）的一个吻。执行一个脚趾，或尾巴捏以确认镇静。如果有必要，麻醉水平，可以进行调整，以获得目标心率为450 ± 50每分钟节拍数（BPM）。 </li><li>轻轻插入直肠探针（润滑油）后，不断监测和调节体温，通过加热垫。重要的是保持在一个狭窄的范围内，体温（37.0 ° C ± 0.5 ° C）时，即使在温度和心率中度变化影响小鼠的心脏功能。 </li><li>四个爪子和磁带他们的心电电极的电极凝胶。 </li></ol> 2。评估心脏收缩功能<ol><li>要开始此过程中，应用超声预热凝胶层到胸部，主要是面积覆心。凝胶应加热到体温近似。冷凝胶的体积可能会迅速增加体温流失率。避免气泡，凝胶，可干扰超声波成像。 </li><li>鼠标固定在小幅上升的位置，下一个很小的角度（抬头）。使用显微操纵器，固定超声探头，探头和心脏之间的90 °角。 </li><li>要开始进行超声心动图测量短轴，降低到胸部parasternally探头。避免直接放置在胸骨或在皮肤上的探头，因为这将扭曲的信号。 </li><li>首先，我们将进行二维（2D）成像（“B型”）获得沿胸骨旁短轴。调整探头的可视面积，以获得一个完整的图像，根据动物的心脏的大小。 </li><li>在这个方向的一个正确的图像将包括左心室和右心室壁略有部分。箭头（视频显示）表明前外侧和后内侧乳头肌，后壁和前壁的LV，脑室室间隔和右心室壁略有部分。在这些测量中，要特别注意潜在的室壁运动异常（例如，运动功能减退，akinasia，异步）或动脉瘤。 </li><li>下一步，我们将使用M型超声心动图，它提供了一个一维（1D）的观点，获得精细测量心脏的尺寸和收缩。商店收购后收缩期左心室功能参数评价的图像。 </li></ol> 3。心脏舒张功能的评价<ol><li>虽然仍查看胸骨旁短轴，我们可以执行组织多普勒成像（TDI）测量心肌运动速度。感兴趣的区域标记，包括后左室壁径向轴评价。得到的波形将有四座山峰评估舒张功能：IVRT，E，A“，并IVCT。 S“代表收缩速度。 （图3B），这些参数将在后面介绍，在代表性的结果部分。 </li><li>可以用脉冲多普勒（PWD）在心肌组织的具体深度来衡量一个很小的范围内血流速度。图像跨二尖瓣血流模式，向后倾斜的动物，在头低脚高的位置。向上倾斜探头，探头正交心尖。 </li><li>现在，增加异氟醚的水平，以降低心率300-350 BPM，这将缓慢运动二尖瓣。比较研究，重要的是要保持动物之间的相似心率。 </li><li>可视化作为参考点，通过阀门的流量测量二尖瓣运动。放置在二尖瓣尖的样本量。 </li><li>测量二尖瓣阀门使用pwd的流动模式。是典型的两波，代表心室的被动填充（早[电子]波），并一个一贯积极填补心房收缩（房[]波）。在一个健康的心脏，E波的速度是比A波（图3C）。 </li><li>一旦完成所有的测量，取出电极凝胶（与标准的纱布轻轻擦拭），从动物的限制。允许奔放的动物在直立位置，并关闭麻醉恢复加热心电图垫之上。当鼠标唤醒，返回到笼子里。 </li></ol> 4。代表经胸超声心动图结果可用于无创性超声心动图研究评估左心室形态和功能在心脏疾病的小鼠模型 ，4,5这项技术被广泛使用，并提供终端程序的替代评估左室功能，如腔内压力-容积循环measurements.8或体外工作心脏模型。LV室尺寸的评估允许经胸M型超声心动图检测异常低压腔扩大或增加患心脏病的各种型号壁厚5因此，经胸超声心动图，结合脉冲多普勒构成的非侵入性的评价，在小鼠心脏收缩和舒张功能的全面大道。 <img src="/files/ftp_upload/1738/1738fig1.jpg" alt="图1" /> 图1。 VEVO 770高分辨率超声心动图和多普勒成像在体内微成像系统概述 （ 一 ）我们用生理监测装置，心电图板，RMV 30MHZ扫描头VisualSonics一体化的铁路系统。 （二）鼠标定位和适当的综合心电电极（金色）加热板的克制。四肢录音到心电图电极。 （ 光盘版 ）用于获取脉冲多普勒适当角度的照片显示，作为在本议定书第4条所述的舒张功能测量。探头收缩功能测量的方向显示在视频协议。 <img src="/files/ftp_upload/1738/1738fig2.jpg" alt="图2" /> 图2。代表二维超声心动图图像（B型）（A）段胸骨短轴显示左心室前壁（AW）和后（星期三）墙壁，室内间隔墙（SW），侧壁（LW）。左心室腔内的直径可以测量左心室内径（LVID）。星号表示后内侧乳头肌。 （二）心尖四腔观，可视面积比测定的E / A峰值速度二尖瓣。 MVAL，二尖瓣前叶; MVPL，二尖瓣后瓣; RV，右心室的LV，左心室。 <img src="/files/ftp_upload/1738/1738fig3.jpg" alt="图3" /> 图3。超声心动图和多普勒数据的采集和定量 。这个数字包含代表图像对应的二维超声心动图图2中的图像采集到的数据。 （ 一 ）M型跟踪线表示收缩末期（ESD）和舒张末直径（EDD）。 LVAWD / s;，左室前壁的厚度（舒张/收缩）LVPWD / S，左室后壁的厚度（舒张/收缩）。 （二）组织多普勒成像的LV后壁代表描。 IVRT，等容舒张时间; IVCT，等容收缩时间。 E“波对应的二尖瓣环的议案在舒张早期充盈的LV，一个”浪潮起源心房收缩后期填写的LV。 S“代表收缩速度。 （c）脉冲多普勒记录二尖瓣单张提示提供二尖瓣血流速度，舒张早期速度（E），舒张收缩与舒张晚期速度（一），E / A比值可以推导出模式。 IVRT也是一个有用的变量来表征舒张功能和充盈压。 ET表示射血时间。

<ol>
	<li>Zhang, T. M. a. i. e. r., S, L., Dalton, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+deltaC+isoform+of+CaMKII+is+activated+in+cardiac+hypertrophy+and+induces+dilated+cardiomyopathy+and+heart+failure.">The deltaC isoform of CaMKII is activated in cardiac hypertrophy and induces dilated cardiomyopathy and heart failure.</a> Circ Res. 92 (8), 912-912 (2003).</li><li>Srivastava, D., Thomas, T., Lin, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cardiac+mesodermal+and+neural+crest+development+by+the+bHLH+transcription+factor,+dHAND.">Regulation of cardiac mesodermal and neural crest development by the bHLH transcription factor, dHAND.</a> Nat Genet. 16 (2), 154-154 (1997).</li><li>Sood, S., Chelu, M. G., van Oort, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+leak+due+to+FKBP12.6+deficiency+in+mice+facilitates+the+inducibility+of+atrial+fibrillation.">Intracellular calcium leak due to FKBP12.6 deficiency in mice facilitates the inducibility of atrial fibrillation.</a> Heart Rhythm. 5 (7), 1047-1047 (2008).</li><li>Gardin, J. M., Siri, F. M., Kitsis, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiographic+assessment+of+left+ventricular+mass+and+systolic+function+in+mice.">Echocardiographic assessment of left ventricular mass and systolic function in mice.</a> Circ Res. 76 (5), 907-907 (1995).</li><li>Tanaka, N., Dalton, N., Mao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transthoracic+echocardiography+in+models+of+cardiac+disease+in+the+mouse.">Transthoracic echocardiography in models of cardiac disease in the mouse.</a> Circulation. 94 (5), 1109-1109 (1996).</li><li>Patten, R. D., Aronovitz, M. J., Bridgman, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+pulse+wave+and+color+flow+Doppler+echocardiography+in+mouse+models+of+human+disease.">Use of pulse wave and color flow Doppler echocardiography in mouse models of human disease.</a> J Am Soc Echocardiogr. 15 (7), 708-708 (2002).</li><li>Garcia-Fernandez, M. A., Azevedo, J., Moreno, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regional+Left+Ventricular+Diastolic+Dysfunction+Evaluated+by+Pulsed-Tissue+Doppler+Echocardiography.">Regional Left Ventricular Diastolic Dysfunction Evaluated by Pulsed-Tissue Doppler Echocardiography.</a> Echocardiography. 16 (5), 491-491 (1999).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nat Protoc. 3 (9), 1422-1422 (2008).</li><li>Larsen, T. S., Belke, D. D., Sas, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pflugers Arch. 437 (6), 979-979 (1999).</li><li>Roth, D. M., Swaney, J. S., Dalton, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+anesthesia+on+cardiac+function+during+echocardiography+in+mice.">Impact of anesthesia on cardiac function during echocardiography in mice.</a> Am J Physiol Heart Circ Physiol. 282 (6), H2134-H2134 (2002).</li></ol>

重要的是要规范麻醉水平，10体温，和心率，促进不同群体之间或基因型的小鼠的比较时使用的M -型超声心动图在小鼠队列。为残疾人士舒张功能的评估，必须降低心率以评价小鼠心脏舒张属性。动物的重要职能，必须密切观察，以防止血液动力学功能衰竭而死亡。另一方面，收缩功能的超声心动图的M -模式，心率每分钟400次以下的测量可能会导致舒张末期直径增加，因为增加填写的LV。

*所需的软件VEVO软件，其中包括B模式的应用，脉冲多普勒应用，和心血管系统的分析工具包。

transthoracic echocardiography in mice

近年来，小鼠模型已成为心功能不全，导致基因表达的变化的分子机制研究的主要途径。转基因和基因打靶的方法，可用于生成与改变的心脏大小和功能的小鼠，<sup&gt; 1-3</sup&gt;作为一个结果，<em&gt;在体内</em&gt;技术需要评估其心脏表型。经胸超声心动图，脉冲多普勒（PWD）和组织多普勒成像（TDI），可用于提供小鼠心脏的三维测量和量化的心脏的收缩和舒张性能程度。二维成像是用来检测异常的解剖或左心室运动，而M型回声是用于心脏的尺寸和收缩的量化。<sup&gt; 4,5</sup此外，PWD是用来量化湍流的本地化速度，<sup&gt; 6</sup&gt;而TDI是用于测量心肌运动速度。<sup&gt; 7</sup因此，经胸超声心动图提供了一个全面的方法在小鼠的心脏功能的无创性评价。

Watch this Scientific Journal Video about Transthoracic Echocardiography in Mice at JoVE.com

Transthoracic Echocardiography in Mice

经胸超声心动图提供了一种无创的检查方法在小鼠心脏功能的评价。超声多普勒成像相结合，可以用来获得三维测量的心和心内的血流量，它们一起提供一个评估心脏的收缩和舒张性能。

经胸超声心动图在小鼠

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

61.5K 次观看.  Baylor College of Medicine (BCM). 经胸超声心动图提供了一种无创的检查方法在小鼠心脏功能的评价。超声多普勒成像相结合，可以用来获得三维测量的心和心内的血流量，它们一起提供一个评估心脏的收缩和舒张性能。

视频: Transthoracic Echocardiography in Mice

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

小鼠超声心动图和超声成像

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

科研

医学

Murine Fetal Echocardiography

Transgenic mice displaying abnormalities in cardiac development and function represent a powerful tool for the understanding the molecular mechanisms underlying both normal cardiovascular function and the pathophysiological basis of human cardiovascular disease. Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development 1-3. In order to study the role of genetic or pharmacologic alterations in the early development of cardiac function, ultrasound imaging of the live fetus has become an important tool for early recognition of abnormalities and longitudinal follow-up. Noninvasive ultrasound imaging is an ideal method for detecting and studying congenital malformations and the impact on cardiac function prior to death 4. It allows early recognition of abnormalities in the living fetus and the progression of disease can be followed in utero with longitudinal studies 5,6. Until recently, imaging of fetal mouse hearts frequently involved invasive methods. The fetus had to be sacrificed to perform magnetic resonance microscopy and electron microscopy or surgically delivered for transillumination microscopy. An application of high-frequency probes with conventional 2-D and pulsed-wave Doppler imaging has been shown to provide measurements of cardiac contraction and heart rates during embryonic development with databases of normal developmental changes now available 6-10. M-mode imaging further provides important functional data, although, the proper imaging planes are often difficult to obtain. High-frequency ultrasound imaging of the fetus has improved 2-D resolution and can provide excellent information on the early development of cardiac structures 11.

Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development. High-frequency ultrasound imaging has improved 2-D resolution and can provide excellent information on early cardiac development and is an ideal method to detect the impact on cardiac structure and function prior to death.

小鼠胎儿超声心动图

Transgenic mice displaying abnormalities in  cardiac development and function represent a powerful tool for the understanding  the molecular mechanisms ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

心率在小鼠中通过无创M型超声心动图测量子宫内

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

右心室结构和功能的肺动脉缩窄经胸超声小鼠模型的评估

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. Experiments involving myocardial injection are currently performed by direct surgical access through a thoracotomy. While convenient when performed at the time of another experimental manipulation such as coronary artery ligation, the need for an invasive procedure for intramyocardial delivery limits potential experimental designs. With ever improving ultrasound resolution and advanced noninvasive imaging modalities, it is now feasible to routinely perform ultrasound-guided, percutaneous intramyocardial injection. This modality efficiently and reliably delivers agents to a targeted region of myocardium. Advantages of this technique include the avoidance of surgical morbidity, the facility to target regions of myocardium selectively under ultrasound guidance, and the opportunity to deliver injectate to the myocardium at multiple, predetermined time intervals. With practiced technique, complications from intramyocardial injection are rare, and mice quickly return to normal activity on recovery from anesthetic. Following the steps outlined in this protocol, the operator with basic echocardiography experience can quickly become competent in this versatile, minimally invasive technique.

Echocardiography-guided percutaneous intramyocardial injection represents an efficient, reliable, and targetable modality for the delivery of gene transfer agents or cells into the murine heart. Following the steps outlined in this protocol, the operator can quickly become competent in this versatile, minimally invasive technique.

超声引导下经皮内注射小鼠

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the examiners&#39; expertise. Speckle tracking Echocardiography (STE) is a promising but technically challenging method that can be used to quantitatively assess regional and global systolic and diastolic myocardial performance. Myocardial strain and strain rate can be measured in all three dimensions&#160;&#8212; radial, circumferential, longitudinal &#8212; of myocardial deformation. Standard cross-sectional two-dimensional B-mode images are recorded and subsequently postprocessed by automated continuous frame-by-frame tracking and motion analysis of speckles within the myocardium. Images are recorded as digital loops and synchronized to a 3-lead EKG for timing purposes. Longitudinal deformation is assessed in the apical 4-, 3-, and 2-chamber views. Circumferential and radial deformation are measured in the parasternal short axis plane.
Optimal image quality and accurate tissue tracking are paramount for the correct determination of myocardial performance parameters. Utilizing transthoracic STE in a healthy volunteer, the present article is a detailed outline of the essential steps and potential pitfalls of quantitative echocardiographic myocardial deformation analysis.

Speckle tracking echocardiography is an emerging diagnostic imaging technique for the quantitative assessment of global and regional myocardial performance. Standard view echocardiographic motion images are recorded and deformation parameters are subsequently measured by automated continuous frame-by-frame tracking and motion analysis of speckles within the B-mode images of the myocardium.

经胸斑点跟踪超声心动图左室心肌变形的定量评价

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic pressure through echocardiography, as well as to obtain measurements of the Aorta and PA diameter in mice.
Mice older than 10 d of age can be analyzed using the present technique. The technique is composed of 3 main steps: set-up, image acquisition, and image analysis. The set-up step consists of getting the mouse anesthetized with 1% isoflurane, shaving it, and taping it in a supine position to a heated EKG board where the image acquisition will take place. The image acquisition step consists of learning to identify the cardiac structures and obtaining all the required images with its correspondent probe and axis in order to be able to calculate volumes and diameters. The image analysis step consists of measuring the previously acquired images with the aid of computer software.
Advantages of the proposed technique include a fast (15 min) procedure that would allow the researcher to evaluate interventions in a non-invasive, non-terminal approach and therefore follow the same mouse over time; each mouse can be used as its own control. This fact plus having the same operator perform all the acquisition and analysis for the entire experiment minimizes the limitation of operator-dependency. The present methodology is useful for mouse researchers in cardiovascular and pulmonary medicine.

The following protocol describes the methodology for the acquisition and analysis of echocardiographic images used to obtain the Left Atrial Volume (LAV), Aorta (Ao) diameter, and Pulmonary Artery (PA) diameter in mice. This technique is a non-invasive, non-terminal procedure that allows assessment of the cardiopulmonary function.

左心房，主动脉和肺动脉的超声心动图小鼠

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

2D and 3D Echocardiography in the Axolotl (Ambystoma Mexicanum)

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model species such as zebrafish and salamanders that are capable of intrinsic heart regeneration hold promise for future regenerative therapies for human patients. To evaluate the outcome of cardioregenerative experiments it is imperative that heart function can be monitored. The axolotl salamander (A. mexicanum) represents a well-established model species in regenerative biology attaining sizes that allows for evaluation of cardiac function. The purpose of this protocol is to establish methods to reproducibly measure cardiac function in the axolotl using echocardiography. The application of different anesthetics (benzocaine, MS-222, and propofol) is demonstrated, and the acquisition of two-dimensional (2D) echocardiographic data in both anesthetized and unanesthetized axolotls is described. 2D echocardiography of the three-dimensional (3D) heart can suffer from imprecision and subjectivity of measurements, and to alleviate this phenomenon a solid method, namely intra/inter-operator/observer analysis, to measure and minimize this bias is demonstrated. Finally, a method to acquire 3D echocardiographic data of the beating axolotl heart at a very high spatiotemporal resolution and with pronounced blood-to-tissue contrast is described. Overall, this protocol should provide the necessary methods to evaluate cardiac function and model anatomy, and flow dynamics in the axolotl using ultrasound imaging with applications in both regenerative biology and general physiological experiments.

2D and 3D Echocardiography in the Axolotl Ambystoma Mexicanum

Here we present echocardiography protocols for two-dimensional and three-dimensional image acquisition of the beating heart of the axolotl salamander (Ambystoma mexicanum), a model species in heart regeneration. These methods allow for longitudinal evaluation of cardiac function at a high spatiotemporal resolution.

axolotl 中的二维和三维超声心动图

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or excision. Many of these nodules are identified and excised via minimally invasive thoracic surgery; however, subcentimeter and subsolid nodules are frequently difficult to identify intra-operatively. This can be mitigated by the use of electromagnetic transthoracic needle localization. This protocol delineates the step-by-step process of electromagnetic localization from the pre-operative period to the postoperative period and is an adaptation of the electromagnetically guided percutaneous biopsy previously described by Arias et al. Pre-operative steps include obtaining a same day CT followed by the generation of a three-dimensional virtual map of the lung. From this map, the target lesion(s) and an entry site are chosen. In the operating room, the virtual reconstruction of the lung is then calibrated with the patient and the electromagnetic navigation platform. The patient is then sedated, intubated, and placed in the lateral decubitus position. Using a sterile technique and visualization from multiple views, the needle is inserted into the chest wall at the prechosen skin entry site and driven down to the target lesion. Dye is then injected into the lesion and, then, continuously during needle withdrawal, creating a tract for visualization intra-operatively. This method has many potential benefits when compared to the CT-guided localization, including a decreased radiation exposure and decreased time between the dye injection and the surgery. Dye diffusion from the pathway occurs over time, thereby limiting intra-operative nodule identification. By decreasing the time to surgery, there is a decrease in wait time for the patient, and less time for dye diffusion to occur, resulting in an improvement in nodule localization. When compared to electromagnetic bronchoscopy, airway architecture is no longer a limitation as the target nodule is accessed via a transparenchymal approach. Details of this procedure are described in a step-by-step fashion.

Presented here is a protocol for lung nodule localization using dye marking via electromagnetically navigated transthoracic needle access. The technique described here can be accomplished in the peri-operative period to optimize nodule localization and to successful resection when performing minimally invasive thoracic surgery.