1。胎盘的制备<ol><li>收购胎盘应尽快从产房分娩后，以尽量减少血凝块形成，干燥和组织的整体恶化。 </li><li>应谨慎，以确保胎盘是不暴露于福尔马林或其他固定液。 </li><li>标准BSL2应注意保持人体组织和​​人员应戴上口罩，隔离衣，手套和鞋套。 </li><li>放置在一个大的塑料或金属碗，预切孔，以便引流液的胎盘。此碗应放在一个较大的不锈钢碗，以防止泄漏。 </li><li>用温生理盐水冲洗胎盘。 </li><li>对于更好胎盘的可视化，消费的跨双羊水膜，确保不损害胎盘本身使用梅岑鲍姆剪刀。留下到位的跨双膜0.5厘米袖口。 （图1A）。 </li></ol> 2。胎盘血管插管<ol><li>确定各自的双胞胎线。产科医生通常每双标识放置在每个线的夹具。请务必确认线提供了展示双（双）。 </li><li>一时间，保持跟踪线，断面每个线正下方的夹子（图1B）。轻轻乳凝块从船只（图1C）。在电源线的切割边缘会有一个静脉和至少一个，通常有两种，动脉。动脉确定其直径相对较小（比静脉）和周围的肌肉袖口（图1D）。 </li><li>使用一个标准的桡动脉行导丝，直径为0.46毫米和80厘米长（箭头，读美国宾夕法尼亚州），用软技巧获得双A的线（图1E）切缘的动脉孔。 </li><li>导丝应放入船只至少3厘米。温和上线使用凯利钳切割边缘的非血管部分柜台紧张，极大地方便了线的进步。线小转向理顺曲折的血管可能有助于用铁丝进步。应谨慎，以避免虚假的段落。 </li><li>一个直言斜面在电线的末端动脉（图1F）使用Seldinger技术，线程20厘米长0.6-1.19毫米内径聚乙烯导管（Intramedic，巴达维亚IL，美国）。必须小心不赶走线同时推进的导管。取下导管后坐在船至少有3厘米的钢丝。 </li><li>另外，某些群体有不使用的脐与脐动脉导管血管的导丝和直接插管成功。虽然，这种技术在我们手中使用的导丝，我们才能完成该过程所花费的时间减少。 </li><li>将一个18 G先端钝针（BD公司，Franlik湖区，新泽西州，美国）油管的近端。确保船只注入少量的生理盐水，观察结算远端动脉导管妥善安置。温柔的手指上的血管注射生理盐水的同时扫描可能有助于促进运动的任何残余的血块。 </li><li>仔细重复步骤2.3至2.6，注意避免撞出双人房A静脉动脉导管。 </li><li>扎脐带1厘米的片，远端切缘周围的电线和导管。确保完全密封，以避免回流。 </li><li>重复步骤2.3至2.9双乙线（非呈现双）等，有脐动脉，静脉和脐磁带和两个环流担保每双导管已用生理盐水（图2）注入。 </li></ol> 3。胎盘血管彩色染料注入<ol><li>病理学染料，钡悬浮液，明胶或其他粘性的彩色染料，可用于可视化的胎盘血管。选择四个对比鲜明的色彩与注入的动脉和静脉对双胞胎一个和B，每双动脉和静脉的颜色应该很容易区分。浅表吻合应染料混合，形成不同（混合）不易察觉的色彩检测。 </li><li>使用3毫升注射器注入动脉双在低压与染料的手。轻轻扫过沿染料流动方向的船只，以确保填充整个血管树与一个或两个手指。如果是确定的，表面的血管吻合手册闭塞饲养容器可以允许较高的抗血管床休息，以填补（图3A）。 </li><li>使用一个单独的手3毫升注射器注入双乙动脉（图3B）。 </li><li>重复步骤3.2和3.3静脉注射。 </li><li>如果受伤的船只，维​​修染料泄漏，应进行罚款缝合针驱动程序。小心不要缝合结扎整个VESSEL。 </li><li>使用一个高分辨率的数码相机，每次注射后的图像。所有注射完成后，取出试样的碗，地方对一个中性色彩背景和重形象（图4）。文件的数量和类型的双胞胎"环流之间看到连接。 </li></ol> 4。代表性的成果: 动脉动脉吻合（AAS）已表明要"保护"，因为它们与一个显著下降TTTS风险和相关的两个双胞胎5改进的成果。图4演示了一个注入MC双胎盘。一节AA前瞻性已确定在子宫内多普勒超声。这一发现证实了混合双人房A动脉（黄色）和双乙动脉（蓝色）在交通动脉结构的染料（绿色箭头）。也表明双人房A（黄色）和双B（橙色）静脉与动脉吻合的AV（箭头）。图5和6描绘我们注入其他两个胎盘标本的照片。 <img src="/files/ftp_upload/2837/2837fig1.jpg" alt="图1"/> 图1。 （一）羊膜小心取出从梅岑鲍姆剪刀共享MC胎盘的胎儿面（B）一对双胞胎的脐带清洁的边缘是由近端切断与梅岑鲍姆剪刀暴露（C ）血块挤奶从人工压迫脊髓（D）脐动脉（箭头）和静脉（箭头）的双A的确定。（e）在放置一个上线的切割面缺血性部分凯利钳牵引，导丝置于脐动脉双A和通过曲折的船只轻轻先进。（F）的小导管，然后在导丝入血管先进，使用Seldinger技术的管腔。 <img src="/files/ftp_upload/2837/2837fig2.jpg" alt="图2"/> 图2。导管已被放置在每个双胞胎中的一个脐动脉和脐静脉。这些导管担保围绕整个线的脐带磁带关系的地方。每个导管的正确定位，已被证实已回暖生理盐水注射清除腔内的血块。 <img src="/files/ftp_upload/2837/2837fig3.jpg" alt="图3"/> 图3。 （一）动脉循环的双胞胎，一个是注射用黄色染料。明显的对比（二）双乙动脉循环是用蓝色染料注入到对双胞胎B.动脉流通传递整个血管赤道。掺加的内表现出的AA吻合（箭头），绿色染料黄色染料和蓝色染料的结果。 <img src="/files/ftp_upload/2837/2837fig4.jpg" alt="图4"/> 图4。静脉双A（红色）和静脉双B（橙色）已注入。机管局吻合（箭头）和一个AV吻合，具有流量指示，从双一对双胞胎乙（箭头）划定。 <img src="/files/ftp_upload/2837/2837fig5.jpg" alt="图5"/> 图5。MC双胎盘接受激光治疗后，产后注射的例子。捐助双胞胎的动脉和静脉分别为黄色和绿色。收件人的动脉和静脉分别为蓝色和橙色。无残留的环流之间的连接进行检测。 <img src="/files/ftp_upload/2837/2837fig6.jpg" alt="图6"/> 图6。MC双胎盘注射后的范例。左侧双胞胎的动脉和静脉分别为黄色和红色。右路双胞胎的动脉和静脉分别为蓝色和橙色。一节AA（箭头）和VV（箭头），胎盘表面上可见。

<ol>
	<li>Rand, L., Lee, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complicated+monochorionic+twin+pregnancies:+updates+in+fetal+diagnosis+and+treatment.">Complicated monochorionic twin pregnancies: updates in fetal diagnosis and treatment.</a> Clin Perinatol. 36, 417-430 (2009).</li><li>Sebire, N., Talbert, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twin-to-twin+transfusion+syndrome+results+from+dynamic+asymmetrical+reduction+in+placental+anastomoses:+a+hypothesis.">Twin-to-twin transfusion syndrome results from dynamic asymmetrical reduction in placental anastomoses: a hypothesis.</a> in Placenta. , (2001).</li><li>Machin, G. A., Keith, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> An atlas of multiple pregnancy: biology and pathology. , (1999).</li><li>Machin, G. A., Feldstein, V. A., van Gemert, M. J., Keith, L. G., Hecher, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+sonographic+demonstration+of+arterio-venous+anastomosis+in+monochorionic+twin+gestation.">Doppler sonographic demonstration of arterio-venous anastomosis in monochorionic twin gestation.</a> Ultrasound Obstet Gynecol. 16, 214-217 (2000).</li><li>Denbow, M. L., Cox, P., Taylor, M., Hammal, D. M., Fisk, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+angioarchitecture+in+monochorionic+twin+pregnancies:+Relationship+to+fetal+growth,+fetofetal+transfusion+syndrome,+and+pregnancy+outcome.">Placental angioarchitecture in monochorionic twin pregnancies: Relationship to fetal growth, fetofetal transfusion syndrome, and pregnancy outcome.</a> American Journal of Obstetrics and Gynecology. 182, 417-426 (2000).</li><li>Taylor, M. J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+detection+of+arterio-arterial+anastomoses+in+monochorionic+twins:+feasibility+and+clinical+application.">Doppler detection of arterio-arterial anastomoses in monochorionic twins: feasibility and clinical application.</a> Human Reproduction. 15, 1632-1636 (2000).</li><li>Berm&#250;dez, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+types+and+twin-twin+transfusion+syndrome.">Placental types and twin-twin transfusion syndrome.</a> American Journal of Obstetrics and Gynecology. 187, 489-494 (2002).</li><li>Lewi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intertwin+anastomoses+in+monochorionic+placentas+after+fetoscopic+laser+coagulation+for+twin-to-twin+transfusion+syndrome:+is+there+more+than+meets+the+eye.">Intertwin anastomoses in monochorionic placentas after fetoscopic laser coagulation for twin-to-twin transfusion syndrome: is there more than meets the eye.</a> Am J Obstet Gynecol. 194, 790-795 (2006).</li><li>Wee, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+of+deep+arterio-venous+anastomoses+within+monochorionic+placentae+by+vascular+casting.">Characterisation of deep arterio-venous anastomoses within monochorionic placentae by vascular casting.</a> Placenta. 26, 19-24 (2005).</li><li>Paepe, M. E. D. e., Burke, S., Luks, F. I., Pinar, H., Singer, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demonstration+of+placental+vascular+anatomy+in+monochorionic+twin+gestations.">Demonstration of placental vascular anatomy in monochorionic twin gestations.</a> Pediatr Dev Pathol. 5, 37-44 (2002).</li></ol>

没有利益冲突的声明。

染料注射MC双胎盘血管的映射是评估复杂，临床治疗的情况下，为进一步加强我们的了解MC的孪生的宝贵的过程。与产科队伍，以确保迅速获得一个完整的胎盘标本的协调是至关重要的。干燥或组织的长期存储在注射之前，可能会导致血管损伤和染料外渗。必须避免胎盘固定，因为交联的血块会阻塞血管染料灌注。在此过程中的关键和困难的一步是脐动脉和静脉插管。这些船只，特别是动脉，是脆...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td>多孔塑料碗</td><td> QuickMedical的医疗设备和用品</td><td> Polarware PA75 </td><td>穿孔，可一钻</td></tr><tr><td>不锈钢碗</td><td>纳朗医药有限公司</td><td> HH517 </td><td></td></tr><tr><td>生理盐水</td><td>费舍尔</td><td> 50983204 </td><td></td></tr><tr><td> 0.6-1.9 mm内径聚乙烯管材</td><td> Intramedic </td><td> 7436 </td><td></td></tr><tr><td>动脉线导丝（0.46毫米） </td><td> arrow国际</td><td> PW - 18080 </td><td>轻柔的笔尖</td></tr><tr><td> 18 G钝性针</td><td> BD公司</td><td> 305180 </td><td></td></tr><tr><td>脐磁带</td><td>质量控制供应</td><td> 140895 </td><td></td></tr><tr><td>梅岑鲍姆剪刀</td><td>科尔 - 帕默</td><td> EW - 10921 - 04 </td><td></td></tr><tr><td>凯利钳</td><td>科尔帕默</td><td> EW - 10918 - 20 </td><td></td></tr><tr><td> 3 ml注射器</td><td> Covidien公司</td><td> 1180300555 </td><td></td></tr><tr><td>病理染料</td><td> Bradley产品</td><td> 7365 - X </td><td></td></tr><tr><td>数码相机</td><td>松下</td><td> LUMIX - TZ50 </td><td></td></tr></tbody></table>

guide wire assisted catheterization and colored dye injection for vascular mapping of monochorionic twin placentas

单绒毛膜双胎妊娠（MC）的是与显着较高的发病率和死亡率比dichorionic双胞胎。大约50％的MC双胎妊娠并发症所产生的共享胎盘及相关血管连接1。严重的双对双胞胎综合征（TTTS）报道，占约20％，这些并发症 2,3 。双胞胎间的血管连接出现在几乎所有的MC胎盘，都涉及到这些高风险的双胎妊娠的预后和结果。 TTTS和其他的MC双条件的发展有牵连的数量，规模和类型的连接。三种类型的双胞胎间的血管连接出现:1）动脉，静脉连接（AVS），其中一个分支动脉在进行无氧血液从胎盘和潜水成胎盘子叶的胎儿面沿双课程。血液流动通过深部脑实质成出现在胎盘的胎儿面，并带来对其他双含氧血的引流静脉的毛细血管网。是从双单向流，提供对接收传出静脉双的入球小动脉; 2）动脉动脉连接（AAS），其中从每双支动脉符合上肤浅胎盘的表面直接导致血管搏动性双向流，3）静脉静脉连接（VVS），在从每双分支静脉符合直接允许低压双向流动的表面上肤浅的胎盘在产科超声检查宫内有针对性的多普勒审讯已被用来识别存在的AV机管局连接4。产前检测产后胎盘注射液的研究已证实的氨基酸都被证明是改善预后相关的两个双胞胎 5 。此外，双胞胎间的血管连接共享胎盘的胎儿面fetoscopic激光烧蚀现在是年初，严重TTTS的首选治疗方法。 产后胎盘注射液的研究提供了有价值的方法，以确认产前多普勒超声检查结果的准确性和胎儿激光治疗的疗效6。单独使用彩色染料手工注入动脉和静脉循环，每双的技术亮点和描绘AVS，AAS，VVS。这个MC的胎盘血管解剖明确的示范，届时可能与多普勒超声检查和新生儿的结果，以提高我们的MC的孪生及其后遗症的病理生理的认识。在这里，我们展示我们的胎盘注射技术。

Watch this Scientific Journal Video about Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas at JoVE.com

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

单绒毛膜（MC）后出生的双胞胎的胎盘血管的映射提供了一个双胞胎"环流之间的血管连接的详细示范的手段。这些连接的不平衡被认为是在MC的孪生并发症，包括双对双胞胎输血症候群的发展发挥了举足轻重的作用。

导丝辅助导管和彩色染料注射的双人单绒毛膜胎盘血管映射

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

guide-wire-assisted-catheterization-colored-dye-injection-for

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

14.8K 次观看.  University of California, San Francisco. 单绒毛膜（MC）后出生的双胞胎的胎盘血管的映射提供了一个双胞胎"环流之间的血管连接的详细示范的手段。这些连接的不平衡被认为是在MC的孪生并发症，包括双对双胞胎输血症候群的发展发挥了举足轻重的作用。

视频: Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Dual Labeling of Neural Crest Cells and Blood Vessels Within Chicken Embryos Using ChickGFP Neural Tube Grafting and Carbocyanine Dye DiI Injection

All developing organs need to be connected to both the nervous system (for sensory and motor control) as well as the vascular system (for gas exchange, fluid and nutrient supply). Consequently both the nervous and vascular systems develop alongside each other and share striking similarities in their branching architecture. Here we report embryonic manipulations that allow us to study the simultaneous development of neural crest-derived nervous tissue (in this case the enteric nervous system), and the vascular system. This is achieved by generating chicken chimeras via transplantation of discrete segments of the neural tube, and associated neural crest, combined with vascular DiI injection in the same embryo. Our method uses transgenic chickGFP embryos for intraspecies grafting, making the transplant technique more powerful than the classical quail-chick interspecies grafting protocol used with great effect since the 1970s. ChickGFP-chick intraspecies grafting facilitates imaging of transplanted cells and their projections in intact tissues, and eliminates any potential bias in cell development linked to species differences. This method takes full advantage of the ease of access of the avian embryo (compared with other vertebrate embryos) to study the co-development of the enteric nervous system and the vascular system.

Dual Labeling of Neural Crest Cells and Blood Vessels Within Chicken Embryos Using ChickGFP Neural Tube Grafting and Carbocyanine Dye DiI Injection

Here we report dual labeling of neural crest cells and blood vessels using chickGFP neural tube intraspecies grafting combined with intra-vascular DiI injection. This experimental technique allows us to simultaneously visualize and study development of the NCC-derived (enteric) nervous system and the vascular system, during organogenesis.

使用小鸡GFP 神经管移植和碳水化合物染料 DiI 注射，在鸡胚胎内对神经囊细胞和血管进行双重标记

All developing organs need to be connected to both the nervous system (for sensory and motor control) as well as the vascular system (for gas exchange, ...

科研

发育生物学

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia polycythemia sequence (TAPS)1,2. Injection studies of twin placentas have shown that such anastomoses are almost invariably present in monochorionic twins and extremely rare in dichorionic twins1. Three types of anastomoses have been documented: from artery to artery, from vein to vein and from artery to vein. Arterio-venous (AV) anastomoses are unidirectional and are referred to as "deep" anastomoses since they proceed through a shared placental cotyledon, whereas arterio-arterial (AA) and veno-venous (VV) anastomoses are bi-directional and are referred to as "superficial" since they lie on the chorionic plate. Both TTTS and TAPS are caused by net imbalance of blood flow between the twins due to AV anastomoses. Blood from one twin (the donor) is pumped through an artery into the shared placental cotyledon and then drained through a vein into the circulation of the other twin (the recipient). Unless blood is pumped back from the recipient to the donor through oppositely directed deep AV anastomoses or through superficial anastomoses, an imbalance of blood volumes occurs, gradually leading to the development of TTTS or TAPS. The presence of an AA anastomosis has been shown to protect against the development of TTTS and TAPS by compensating for the circulatory imbalance caused by the uni-directional AV anastomoses1,2. 
Injection of monochorionic placentas soon after birth is a useful mean to understand the etiology of various (hematological) complications in monochorionic twins and is a required test to reach the diagnosis of TAPS2. In addition, injection of TTTS placentas treated with fetoscopic laser surgery allows identification of possible residual anastomoses3-5. This additional information is of paramount importance for all perinatologists involved in the management and care of monochorionic twins with TTTS or TAPS. Several placental injection techniques are currently being used. We provide a simple protocol to accurately evaluate the presence of (residual) vascular anastomoses using colored dye injection.

Twin-to-twin transfusion syndrome and twin anemia polycythemia sequence are two potentially devastating problems in perinatal medicine. Both disorders occur only in monochorionic twins and result from unbalanced blood flow through placental vascular anastomoses. We provide a simple protocol to accurately evaluate the presence of vascular anastomoses using colored dye injection of placental vessels after birth.

在单绒毛膜胎盘血管吻合用有色染料的精确和简单的评价

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

医学

Visualization and Quantitative Analysis of Embryonic Angiogenesis in Xenopus tropicalis

Blood vessels supply oxygen and nutrients throughout the body, and the formation of the vascular network is under tight developmental control. The efficient in vivo visualization of blood vessels and the reliable quantification of their complexity are key to understanding the biology and disease of the vascular network. Here, we provide a detailed method to visualize blood vessels with a commercially available fluorescent dye, human plasma acetylated low density lipoprotein DiI complex (DiI-AcLDL), and to quantify their complexity in Xenopus tropicalis. Blood vessels can be labeled by a simple injection of DiI-AcLDL into the beating heart of an embryo, and blood vessels in the entire embryo can be imaged in live or fixed embryos. Combined with gene perturbation by the targeted microinjection of nucleic acids and/or the bath application of pharmacological reagents, the roles of a gene or of a signaling pathway on vascular development can be investigated within one week without resorting to sophisticated genetically engineered animals. Because of the well-defined venous system of Xenopus and its stereotypic angiogenesis, the sprouting of pre-existing vessels, vessel complexity can be quantified efficiently after perturbation experiments. This relatively simple protocol should serve as an easily accessible tool in diverse fields of cardiovascular research.

Visualization and Quantitative Analysis of Embryonic Angiogenesis in Xenopus tropicalis

This protocol demonstrates a fluorescence-based method to visualize the vasculature and to quantify its complexity in Xenopus tropicalis. Blood vessels can be imaged minutes after the injection of a fluorescent dye into the beating heart of an embryo after genetic and/or pharmacological manipulations to study cardiovascular development in vivo.

胚胎血管生成的可视化和定量分析<em&gt;热带非洲爪蟾</em

Blood vessels supply oxygen and nutrients throughout the body, and the formation of the vascular network is under tight developmental control. The ...

Intravascular Perfusion of Carbon Black Ink Allows Reliable Visualization of Cerebral Vessels

The anatomical structure of cerebral vessels is a key determinant for brain hemodynamics as well as the severity of injury following ischemic insults. The cerebral vasculature dynamically responds to various pathophysiological states and it exhibits considerable differences between strains and under conditions of genetic manipulations. Essentially, a reliable technique for intracranial vessel staining is essential in order to study the pathogenesis of ischemic stroke. Until recently, a set of different techniques has been employed to visualize the cerebral vasculature including injection of low viscosity resin, araldite F, gelatin mixed with various dyes1 (i.e. carmine red, India ink) or latex with2 or without3 carbon black. Perfusion of white latex compound through the ascending aorta has been first reported by Coyle and Jokelainen3. Maeda et al.2 have modified the protocol by adding carbon black ink to the latex compound for improved contrast visualization of the vessels after saline perfusion of the brain. However, inefficient perfusion and inadequate filling of the vessels are frequently experienced due to high viscosity of the latex compound4. Therefore, we have described a simple and cost-effective technique using a mixture of two commercially available carbon black inks (CB1 and CB2) to visualize the cerebral vasculature in a reproducible manner5. We have shown that perfusion with CB1+CB2 in mice results in staining of significantly smaller cerebral vessels at a higher density in comparison to latex perfusion5. Here, we describe our protocol to identify the anastomotic points between the anterior (ACA) and middle cerebral arteries (MCA) to study vessel variations in mice with different genetic backgrounds. Finally, we demonstrate the feasibility of our technique in a transient focal cerebral ischemia model in mice by combining CB1+CB2-mediated vessel staining with TTC staining in various degrees of ischemic injuries.

Analysis of rodent cerebrovascular anatomy plays an important role in experimental stroke research. In this context, intravascular perfusion with colored latex has been considered as a standard tool for several years. However, this technique implies distinct technical limitations, which undermine its reproducibility. Here, we describe a simple method to visualize cerebral vessels in a reproducible manner. Injection of a mixture of two commercially available carbon black inks through the left myocardial ventricle results in adequate filling of cerebral vessels with high contrast visualization. We have successfully applied this technique to identify anastomotic points between cerebral vascular territories of mice with different genetic backgrounds. We finally give evidence that this novel and simple method for vessel staining can be combined with triphenyltetrazolium chloride (TTC) staining - a widely used tool to observe and analyze infarct volumes in mice.

血管内灌注的碳黑色墨水可实现可靠的可视化脑血管

The anatomical structure of cerebral vessels is a key determinant for brain hemodynamics as well as the severity of injury following ischemic insults. The ...

神经科学

Ex Vivo Perfusion of the Rodent Placenta

The placenta is a key organ during pregnancy that serves as a barrier to fetal xenobiotic exposure and mediates the exchange of nutrients for waste. An assay is described here to perfuse an isolated rat placenta and evaluate the maternal-to-fetal translocation of xenobiotics ex vivo. In addition, the evaluation of physiological processes such as fluid flow to the fetus and placental metabolism may be conducted with this methodology. This technique is suitable for evaluating maternal-to-fetal kinetics of pharmaceutical candidates or environmental contaminants. In contrast to current alternative approaches, this methodology allows the evaluation of the isolated maternal-fetal vasculature, with the systemic neural or immune involvement removed, allowing any observed changes in physiological function to be attributable to local factors within the isolated tissue.

Here is presented a protocol of ex vivo maternal-fetal vascular perfusion to enable the administration of a test article into maternal vasculature and to evaluate placental transfer of xenobiotic particles or pharmacological agents in addition to alterations in placental physiology.

啮齿类动物胎盘的体外灌注

The placenta is a key organ during pregnancy that serves as a barrier to fetal xenobiotic exposure and mediates the exchange of nutrients for waste. An ...

Non-fluoroscopic Catheter Tracking for Fluoroscopy Reduction in Interventional Electrophysiology

A technological platform (MediGuide) has been recently introduced for non-fluoroscopic catheter tracking. In several studies, we have demonstrated that the application of this non-fluoroscopic catheter visualization system (NFCV) reduces fluoroscopy time and dose by 90-95% in a variety of electrophysiology (EP) procedures. This can be of relevance not only to the patients, but also to the nurses and physicians working in the EP lab. Furthermore, in a subset of indications such as supraventricular tachycardias, NFCV enables a fully non-fluoroscopic procedure and allows the lab staff to work without wearing lead aprons. With this protocol, we demonstrate that even complex procedures such as ablations of atrial fibrillation, that are typically associated with fluoroscopy times of &#62;30 min in conventional settings, can safely be performed with a reduction of &#62;90% in fluoroscopy exposure by the additional use of NFCV.

Radiation exposure is an underestimated risk in complex ablation procedures. Here, we describe a protocol to significantly decrease fluoroscopy time and dosage for both the patient and the lab staff by using a novel non-fluoroscopic catheter visualization system.

非透视导管跟踪透视减少介入电

A technological platform (MediGuide) has been recently introduced for non-fluoroscopic catheter tracking. In several studies, we have demonstrated that ...

An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury

Spinal cord injury (SCI) causes significant vascular disruption at the site of injury. Vascular pathology occurs immediately after SCI and continues throughout the acute injury phase. In fact, endothelial cells appear to be the first to die after a contusive SCI. The early vascular events, including increased permeability of the blood-spinal cord barrier (BSCB), induce vasogenic edema and contribute to detrimental secondary injury events caused by complex injury mechanisms. Targeting the vascular disruption, therefore, could be a key strategy to reduce secondary injury cascades that contribute to histological and functional impairments after SCI. Previous studies were mostly performed on postmortem samples and were unable to capture the dynamic changes of the vascular network. In this study, we have developed an in vivo duo-color two-photon imaging method to monitor acute vascular dynamic changes following contusive SCI. This approach allows detecting blood flow, vessel diameter, and other vascular pathologies at various sites of the same rat pre- and post-injury. Overall, this method provides an excellent venue for investigating vascular dynamics.

An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury

We introduce an in vivo imaging method using two different fluorescent dyes to track dynamic spinal vascular changes following a contusive spinal cord injury in adult Sprague-Dawley rats.

在体内挫伤脊髓损伤后影像血管动力学的双色法

Spinal cord injury (SCI) causes significant vascular disruption at the site of injury. Vascular pathology occurs immediately after SCI and continues ...

Ablation of Ischemic Ventricular Tachycardia Using a Multipolar Catheter and 3-dimensional Mapping System for High-density Electro-anatomical Reconstruction

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars represent zones of slow conduction that allow the occurrence and maintenance of reentrant circuits. Catheter ablation enables substrate modification of those low voltage areas and thus can help to alter the scar tissue in such a way that arrhythmias cannot appear anymore. Hospitalizations of concerned patients decrease, quality of life and outcome rise. Consequently, VT ablation represents a growing field in electrophysiology, especially for patients with endocardial scars in ischemic heart disease after myocardial infarction. However, ablation of ventricular tachycardia remains one of the most challenging procedures in the electrophysiology lab. Precise scar definition and localization of abnormal potentials are critical for ablation success. The following manuscript describes the use of a multipolar mapping catheter and 3-dimensional (3D) mapping system to create a high density electro-anatomical map of the left ventricle including a precise scar representation as well as mapping of fractionated and late potentials in order to allow a highly accurate substrate modification.

With the following protocol, we provide an approach to Ventricular Tachycardia (VT) ablation using high density mapping with a multipolar catheter and 3D mapping system enhancing the success of the procedure.

采用多极导管三维测图系统进行缺血性室性心动过速的高密度电解剖重建

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars ...

Model Surgical Training: Skills Acquisition in Fetoscopic Laser Photocoagulation of Monochorionic Diamniotic Twin Placenta Using Realistic Simulators

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome (TTTS), but is technically challenging and can lead to significant complications. Acquiring and maintaining the necessary surgical skills require consistent practice, a critical caseload, and time. Training on realistic surgical simulators can potentially shorten this steep learning curve and enables several proceduralists to acquire procedure-specific skills simultaneously. Here we describe realistic simulators designed to allow the user familiarity with the equipment and specific steps required in the surgical treatment of TTTS, including fetoscopic handling, approaches to anterior and posterior placenta, recognition of anastomoses, and efficient coagulation of vessels. We describe the skills that are especially important in conducting placental laser coagulation that the surgeon can practice on the model and apply in a clinical case. These models can be adapted easily depending on the availability of materials and require standard fetoscopy equipment. Such training systems are complementary to traditional surgical apprenticeships and can be useful aids for fetal medicine units that provide this clinical service.

Practicing the specific skills required for fetoscopic laser coagulation of monochorionic placental anastomoses on realistic models can aid less experienced surgeons in overcoming the steep learning curve associated with this procedure that is now regarded as the standard of care for twin-twin transfusion syndrome.

模型外科训练: 用逼真模拟器 Fetoscopic 激光光凝单绒毛膜 Diamniotic 双胎胎盘的技能习得

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome ...

Evaluation of Color Difference in Placenta with Twin Anemia Polycythemia Sequence

Twin anemia polycythemia sequence (TAPS) occurs in 5% of monochorionic twins and is characterized by large inter-twin hemoglobin differences. The postnatal diagnostic criteria for TAPS are based on hematologic parameters and placental characteristics. Placental examination after birth shows that color of the maternal side between placental territories of the anemic and polycythemic twins is remarkably different. The color difference in TAPS placentas is higher compared to monochorionic placentas with acute peripartum feto-fetal transfusion; thus, this is used as an additional diagnostic criterion for TAPS. Software such as ImageJ enables the computer-based measurement of color intensity in TAPS placentas. However, a detailed method for the calculation of color differences between anemic and polycythemic components of TAPS placentas has not yet been described. The protocol presented here provides a step-by-step method for analyzing color differences in the maternal side of TAPS placentas using ImageJ software.

In monochorionic twins with twin anemia polycythemia sequence (TAPS), the donor twin and its corresponding placenta share are pale, while the recipient twin and its placental share have a plethoric aspect. Presented here is a protocol to quantify the color difference in maternal side of TAPS placenta after birth.

导丝辅助导管和彩色染料注射的双人单绒毛膜胎盘血管映射

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使用小鸡^GFP神经管移植和碳水化合物染料 DiI 注射，在鸡胚胎内对神经囊细胞和血管进行双重标记