Name: high frequency ultrasound for the analysis of fetal and placental development in vivo
Uploaded: 2018-11-08T14:30:00
Description: Watch this Scientific Journal Video about High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo at JoVE.com

非常感谢成像仪器公司 (特别是 magdalena steiner、katin suppelt 和 sandra meyer) 提供了令人愉快和快速的支持, 并及时、全面地回答了我们所有有关成像系统及其使用的问题。我们感谢汉斯-雷默·罗德瓦尔德教授和索尔斯滕·费耶拉本德博士 (德国德凯斯兹·海德尔贝格) 提供了 cpa3 殖民地。此外, 我们还要感谢 stefanie langwisch, 他负责鼠标群, 并生成了图1中的图片。
这项工作和成像系统的资金来自德国防方大学 (dfg) 至 a. c. z. (ze526x-1 和 az526/1) 的赠款, 这些项目是 dfg 1394 "健康和疾病中的肥大细胞" 中的项目。

这里描述的所有方法都已得到 "landesverwaltungsamt sachsen anstel:42502-2-1296unimd" 的批准。
1. 实验程序
<ol><li>将6至8周龄女性 mc 缺乏 c57bl/6j-cpa3cre/+ (cpa3cre/+) 小鼠和 mcc-充足的 c57bl/6j-cpa3++ (菌落对照;cpa3+/+) 与 balb1 男性。</li>
	<li>定义妊娠日 (gestation) 0 后确认阴道塞和治疗女性后立即塞确认。 注: 塞?...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+sonar+receiver+of+the+bat.">The sonar receiver of the bat.</a> Annals of the New York Academy of Sciences. 188, 161-174 (1971).</li><li>Zala, S. M., Reitschmidt, D., Noll, A., Balazs, P., Penn, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex-dependent+modulation+of+ultrasonic+vocalizations+in+house+mice+(Mus+musculus+musculus).">Sex-dependent modulation of ultrasonic vocalizations in house mice (Mus musculus musculus).</a> Public Library of Science ONE. 12 (12), e0188647 (2017).</li><li>W&#246;hr, M., Seffer, D., Schwarting, R. K. 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B., Ewertsen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonography+of+the+Kidney:+A+Pictorial+Review.">Ultrasonography of the Kidney: A Pictorial Review.</a> Diagnostics. 6 (1), (2015).</li><li>Older, R. A., Watson, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+anatomy+of+the+normal+male+reproductive+tract.">Ultrasound anatomy of the normal male reproductive tract.</a> Journal of Clinical Ultrasound. 24 (8), 389-404 (1996).</li><li>Reeves, J. J., Rantanen, N. W., Hauser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transrectal+real-time+ultrasound+scanning+of+the+cow+reproductive+tract.">Transrectal real-time ultrasound scanning of the cow reproductive tract.</a> Theriogenology. 21 (3), 485-494 (1984).</li><li>Sharma, M., Somani, P., Sunkara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+of+gall+bladder+by+endoscopic+ultrasound.">Imaging of gall bladder by endoscopic ultrasound.</a> World Journal of Gastrointestinal Endoscopy. 10 (1), 10-15 (2018).</li><li>Weskott, H. -. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultraschall+in+der+Diagnostik+maligner+Lymphome.">Ultraschall in der Diagnostik maligner Lymphome.</a> Der Radiologe. 52 (4), 347-359 (2012).</li><li>Shirinifard, A., Thiagarajan, S., Johnson, M. D., Calabrese, C., Sablauer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+Absolute+Blood+Perfusion+in+Mice+Using+Dynamic+Contrast-Enhanced+Ultrasound.">Measuring Absolute Blood Perfusion in Mice Using Dynamic Contrast-Enhanced Ultrasound.</a> Ultrasound in Medicine &amp; Biology. 43 (8), 1628-1638 (2017).</li><li>Quaia, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+tissue+perfusion+by+contrast-enhanced+ultrasound.">Assessment of tissue perfusion by contrast-enhanced ultrasound.</a> European Radiology. 21 (3), 604-615 (2011).</li><li>Saw, S. N., Poh, Y. W., Chia, D., Biswas, A., Zaini Mattar, C. N., Yap, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+hemodynamic+wall+shear+stresses+in+human+umbilical+vessels+from+normal+and+intrauterine+growth+restricted+pregnancies.">Characterization of the hemodynamic wall shear stresses in human umbilical vessels from normal and intrauterine growth restricted pregnancies.</a> Biomechanics and Modeling in Mechanobiology. , (2018).</li><li>Kessler, J., Rasmussen, S., Godfrey, K., Hanson, M., Kiserud, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+growth+restriction+is+associated+with+prioritization+of+umbilical+blood+flow+to+the+left+hepatic+lobe+at+the+expense+of+the+right+lobe.">Fetal growth restriction is associated with prioritization of umbilical blood flow to the left hepatic lobe at the expense of the right lobe.</a> Pediatric Research. 66 (1), 113-117 (2009).</li><li>Laurin, J., Lingman, G., Mars&#225;l, K., Persson, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+blood+flow+in+pregnancies+complicated+by+intrauterine+growth+retardation.">Fetal blood flow in pregnancies complicated by intrauterine growth retardation.</a> Obstetrics and Gynecology. 69 (6), 895-902 (1987).</li><li>Arduini, D., Rizzo, G., Romanini, C., Mancuso, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+blood+flow+velocity+waveforms+as+predictors+of+growth+retardation.">Fetal blood flow velocity waveforms as predictors of growth retardation.</a> Obstetrics and Gynecology. 70 (1), 7-10 (1987).</li><li>Meyer, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chymase-producing+cells+of+the+innate+immune+system+are+required+for+decidual+vascular+remodeling+and+fetal+growth.">Chymase-producing cells of the innate immune system are required for decidual vascular remodeling and fetal growth.</a> Scientific Reports. 7, 45106 (2017).</li><li>Meyer, N., Sch&#252;ler, T., Zenclussen, A. 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利用我们的超声系统, 我们展示了从 gd10 开始缺乏 mcw 的母亲胎儿生长的限制。此外, 在 gd10 和 12, 我们观察到胎盘尺寸减少, 在 gd10 中, 一些缺乏 umcm函的小鼠在 uma 中没有或逆转端部舒张流。这种血管化不良的迹象与动脉的显著阻力指数有关。结果证实了 uMCs 和 uMCs 在妊娠和胎儿健康以及了解 iugr 过程中的重要作用。
该协议适用于从 gd5 开始 (植入后) 的每一个妊娠日。议定书?...

超声波是高于人耳听觉范围的声波, 高于约20千赫 1.像蝙蝠、威尔士、海豚2、3、老鼠4、老鼠5和老鼠狐猴 6这样的动物都使用超声波进行定向或交流。人类利用超声波进行多种技术和医疗应用。超声波装置能够产生声波, 并传播和表示信号。如果超声波遇到障碍, 声音会被反射、吸收或通过。超声作为一种成像方法的应用, 称为超声, 用于分析人体或兽医的有机组织, 如心脏 (超声心动图)7,8, 肺 9, 甲状腺10, 肾脏11, 和尿和生殖道12,13;检测胆结石14和肿瘤15;并评估血管或器官的灌注16,17。超声是怀孕期间产前护理的标准方法, 胎儿发育障碍或损伤可以及早识别。具体而言, 定期密切监测胎儿的生长情况, 以识别可能的 iugr。最后, 可以监测胎儿的血液流动情况, 因为这可以指出生长限制18、19、20、21.
与其他方法 (如放射学) 相比, 超声成像的一个主要优点是要分析的组织的声音无害性。这种简单快捷的方法是无创、无痛的, 可以多次使用。超声波设备的初始支出昂贵;然而, 所需的消耗品是便宜的。本手稿中使用的超声波系统适用于一系列动物模型(即小鼠和鱼类), 而对于人类而言, 超声波设备需要 3-15 mhz 的频率, 小鼠需要 15-70 mhz 的频率。
本文介绍了一种使用 b 模式、彩色多普勒模式和脉冲波多普勒模式的协议。描述包括小鼠的准备以及性能、数据采集和存储。该方法已成功地应用于不同的小鼠菌株在所有妊娠期, 可用于调查胎儿和胎盘发育以及母亲和胎儿的血液参数。在这里, 所有的应用都解释了基于我们的研究使用怀孕的 mc nk 缺乏和控制小鼠。

<table border="0" cellpadding="0" cellspacing="0" width="758">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">LEAF anti-Maus CD122 (IL-2Rb)</td>
			<td style="width:204px;">BioLegend</td>
			<td style="width:151px;">123204</td>
			<td style="width:187px;">Klon TM-&#946;1; 500 &#181;g</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vevo 2100 System</td>
			<td>&#160;FujiFilm VisualSonics Inc.</td>
			<td style="width:151px;"></td>
			<td>Transducer&#160;MS550D-0421</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vevo LAB Software</td>
			<td>&#160;FujiFilm VisualSonics Inc.</td>
			<td style="width:151px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Baxter</td>
			<td>PZN: 6497131</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Electrode gel</td>
			<td>Parker</td>
			<td>12_8</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical tape</td>
			<td>3M Transpore</td>
			<td>1527-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Eye cream</td>
			<td>Bayer</td>
			<td>PZN: 1578675</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotton tipped applicators</td>
			<td>Raucotupf</td>
			<td>11969</td>
			<td>100 pieces</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Depilatory cream</td>
			<td>Reckitt Benckiser</td>
			<td>2077626</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Compresses</td>
			<td>Nobamed Paul Danz AG</td>
			<td>856110</td>
			<td>10 x 10 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ultrasound gel</td>
			<td>Gello GmbH</td>
			<td>246000</td>
			<td></td>
		</tr>
	</tbody>
</table>

high frequency ultrasound for the analysis of fetal and placental development in vivo

超声成像是一种广泛用于检测人体和动物组织器官异常和肿瘤的方法。该方法无创、无害、无痛, 应用简单、快速, 即使使用移动设备, 也可以在任何地方完成。在怀孕期间, 超声成像被用来密切监测胎儿的发育。该技术是重要的评估宫内生长限制 (iugr), 妊娠并发症, 对母亲和胎儿的短期和长期健康的影响。了解 iugr 的过程对于制定有效的治疗策略是不可或缺的。
本手稿中使用的超声波系统是为分析小动物而产生的一种超声波装置, 可用于包括怀孕研究在内的各种研究领域。在这里, 我们描述了系统的使用, 在体内分析胎儿的自然杀手 (nk) 细胞肥大细胞 (mc) 缺乏的母亲, 生的生长限制的幼崽。该协议包括系统的准备、测量前和测量过程中小鼠的处理, 以及 b 模式、彩色多普勒模式和脉冲波多普勒模式的使用。分析了胎儿的大小、胎盘大小和胎儿的血液供应情况。我们发现, 从妊娠中期开始, 缺乏 nkcc 的小鼠的植入面积和胎盘变小。此外, mc/nk 缺乏与胎儿脐叶(uma) 中无端舒张流和反转端舒张流和阻力指数升高有关。协议中描述的方法可以很容易地用于相关和非相关的研究主题。

本手稿中使用的超声波系统的各个组成部分如图 1所示。图 2显示了在 gd5、8、10和 12 (b)和相应的植入面积测量结果(a)以 b 模式获得的代表性超声图像, 显示植入面积显著减少。从 gd10 开始, 抗 gd10 治疗的 cpa3cre/+小鼠。
图 3显示?...

Watch this Scientific Journal Video about High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo at JoVE.com

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

本文介绍了高频超声对小鼠胎儿体内分析的技术。这种方法允许胎儿的随访和胎盘参数的分析, 以及整个怀孕期间的产妇和胎儿血液流动。

高频超声在胎儿和胎盘发育分析中的影响

Ultrasound imaging is a widespread method used to detect organ anomalies and tumors in human and animal tissues. The method is non-invasive, harmless, and ...

这种方法对于回答肿瘤学、神经生物学、心脏病学和胎儿发育领域的关键问题非常重要，当然在生殖研究领域也很重要。这种技术的主要优点是无创，无痛，容易，快速，并允许胎儿在相关的妊娠时间点在整个怀孕期间随和。在成像程序前至少半小时，打开超声波系统、加热平台和凝胶加热器。在浏览器中的现有研究中打开新的研究或新系列。在研究信息窗口中输入所有必需的信息。单击"确定"，并确认出现 B 模式成像窗口。当超声波准备就绪时，确认麻醉小鼠对手趾捏没有反应，并在动物的眼睛上涂抹软膏。将一滴电凝胶放在加热平台的 4 个铜部分上，然后将爪子固定到平台的电凝胶涂层部分。用脱毛霜和棉签从动物腹部去除头发，一分钟后用水浸压缩洗净脱毛皮肤的乳霜。然后将预热超声波凝胶涂抹在外露区域。信号强度低的一个原因可能是鼠标和超声波束之间的凝胶量低。根据我们的经验，大量的凝胶是良好的信号质量所必需的。当动物准备好时，将传感器放在膀胱上作为参考点，将加热平台表移动到腹部的左右两侧，以 2D B 模式跟踪胎儿植入，必要时调整传感器或平台，直到屏幕上显示第一个植入，其最大尺寸。选择"图像标签"，然后输入图像文件的名称。调整图像的位置，直到胎盘脐带内的血流在视图中，并存储一个框架或 cineloop 进行胎盘测量。要进行彩色多普勒成像以确定血流的方向，请按"颜色"按钮，并使用轨迹球将颜色框移动到适当的位置，单击"更新"可根据需要调整血流框的大小。当框大小正确时，单击"选择"并存储图像作为实验适当。对于流经血管的脉冲波多普勒血流，在彩色多普勒采集模式下定位感兴趣的区域，然后单击脉冲波。将出现虚线。将线移动到感兴趣的血管，并使用多普勒角度旋钮调整线的角度，以与血流对齐。对于脉冲波多普勒测量，观察血流方向和超声波束之间的角度非常重要。在同一实验中，高度的角度或动物之间的不同角度可能会导致速度测量不准确。然后，单击"更新"并存储脉冲波多普勒采集窗口中出现多普勒线的 cineloop。若要查看数据，请单击"研究管理"，滚动到感兴趣的缩略图图像，然后双击"更新"，然后再次单击"关闭"以关闭浏览器窗口，以完成数据采集并保存录制的系列。若要导出数据，请连接硬盘，单击"导出到"，以标记一个或多个系列数据，然后选择用于将数据复制到硬盘上的存储空间。在这里，在妊娠日5、8、10和12时在B模式下采集的代表性超声图像，并显示出相应的植入面积测量结果，显示从妊娠日10日起，抗CD122治疗的乳腺细胞缺陷小鼠的植入面积显著减少。B 模式成像有助于成像和测量植入的各个部分。例如，与妊娠日10和12日未经治疗的小鼠相比，经过治疗的乳腺细胞缺陷动物的防CD122乳腺细胞缺陷动物的胎盘面积、厚度和直径显著减少。相比之下，妊娠期第14天的组与地膜厚度相似，与妊娠期第14天未经治疗的小鼠相比，抗CD122治疗的转基因动物显著增加。脉冲波多普勒成像的子宫动脉测量峰值收缩速度和舒张速度，并计算的阻力指数，揭示了治疗动物和未经治疗的动物之间的类似值。然而，与未经治疗的小鼠相比，抗CD122治疗的乳腺细胞缺陷动物的抗药性指标显著增加。在尝试这个程序时，重要的是要记住，你应该每隔一天，而不是每一天这样做，以避免压力，和重复性地对动物的自恋的风险。按照此过程，还可以进行其他测量，如 3D 测量、组织移动的可视化和定量、血压测量以及超声波导注。在它的发展之后，这种方法为妇产科和产前护理领域的医生和研究人员铺平了道路，因为可以跟踪胎儿发育，并在怀孕期间识别可能的异常。

Research

JoVE Journal

Developmental Biology

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

对比成像的小鼠胚胎使用高频超声

Ultrasound contrast-enhanced imaging can convey essential quantitative information regarding tissue vascularity and perfusion and, in targeted ...

科研

发育生物学

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

斑马鱼幼虫骨骼肌诚信与伊文思蓝分析

The zebrafish model is an emerging system for the study of neuromuscular disorders.  In the study of neuromuscular diseases, the integrity of the muscle ...

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

三维高频超声对小鼠早期妊娠特征的新应用

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

Zika Virus Infection of Cultured Human Fetal Brain Neural Stem Cells for Immunocytochemical Analysis

人胎脑神经干细胞 Zika 病毒感染的免疫细胞化学分析

Human fetal brain neural stem cells are a unique non-genetically modified model system to study the impact of various stimuli on human developmental ...

Three-dimensional Rendering and Analysis of Immunolabeled, Clarified Human Placental Villous Vascular Networks

Immunolabeled, 澄清人胎盘长柔毛血管网络的三维渲染与分析

Nutrient and gas exchange between mother and fetus occurs at the interface of the maternal intervillous blood and the vast villous capillary network that ...

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound (30/45MHZ) System

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound 30/45MHZ System

用高频超声 (30/45 mhz) 系统对胎儿小鼠进行心血管成像

Congenital heart defects (CHDs) are the most common cause of childhood morbidity and early mortality. Prenatal detection&#160;of the underlying molecular ...

Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton

Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton

秀丽线虫生殖的计算分析研究细胞核、蛋白质和细胞骨架的分布

The Caenorhabditis elegans (C. elegans) germline is used to study several biologically important processes including stem cell development, apoptosis, and ...

Intravenous and Intra-amniotic In Utero Transplantation in the Murine Model

Intravenous and Intra-amniotic In Utero Transplantation in the Murine Model

小鼠宫内移植中静脉和羊膜内植入的研究

In utero transplantation (IUT) is a unique and versatile mode of therapy that can be used to introduce stem cells, viral vectors, or any other substances ...

High-Frequency Ultrasound Echocardiography to Assess Zebrafish Cardiac Function

高频超声超声心动图评估斑马鱼心脏功能

The zebrafish (Danio rerio) has become a very popular model organism in cardiovascular research, including human cardiac diseases, largely due to its ...

Transuterine Fetal Tracheal Occlusion Model in Mice

小鼠的转宫胎儿气管闭塞模型

Fetal tracheal occlusion (TO), an established treatment modality, promotes fetal lung growth and survival in severe congenital diaphragmatic hernia (CDH). ...