Name: in vivo imaging of the mouse spinal cord using two-photon microscopy
Uploaded: 2012-01-05T13:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy at JoVE.com

1。大厦的脊椎稳定装置<ol><li>订购Narishige STS的紧凑型脊髓夹钳和Narishige马6N头控股适配器。 </li><li>自定义的设计和制造的不锈钢底板举行对齐Narishige的两个部分，使动物的头是支持，而其脊柱和尾巴夹紧。请记住，整个装置应适合在显微镜下的镜头，通常在降低显微镜阶段。 </li></ol> 2。动物手术<ol><li>预热空气加热装置的显微镜室，并保持在37 ° C。 </li><li>称量动物和麻醉?...

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这里介绍的方法， 可以在体内成像人口稠密荧光灯在使用双光子显微镜的麻醉小鼠的脊髓细胞结构稳定和重复性。所取得的稳定，是一个特制的脊椎稳定装置和麻醉方案，减少呼吸道引起的运动伪影的结果。脊椎稳定装置允许下的鼠体呼吸的空间，可使用市售的脊髓夹具和头部安装片（图1）。高重现性好，微创的方法是，始终没有广泛的图像后处理（图和视频1），可用于实验分析产生的?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>若丹明B葡聚糖</td><td> Invitrogen公司</td><td> D1841 </td><td> 70 kDa的，稀释 学联（3％W / V） </td></tr><tr><td>氯胺酮盐酸</td><td> Bionichepharma </td><td> NDC的无：67457-001-10 </td><td>注射剂，50mg/ml </td></tr><tr><td> Anased </td><td>劳埃德实验室</td><td> NADA无：139-236 </td><td>甲苯噻嗪注射剂， 20mg/ml </td></tr><tr><td> Acepromazine </td><td> Vedco </td><td> NADA无：117-531 </td><td>注射剂，10mg/ml的</td></tr><tr><td>人工泪液 软膏</td><td>凤凰 医药</td><td> NDC：57319-760 - 25 </td><td>润滑剂</td></tr><tr><td>优碘</td><td>费舍尔</td><td> 19-061617 </td><td></td></tr><tr><td>麦克弗森的韦斯科特 剪刀</td><td>世界精密 仪器</td><td> 555500S </td><td>弯曲，钝尖 剪刀</td></tr><tr><td>直钳</td><td>世界精密 仪器</td><td> 555047FT </td><td>齿尖钳</td></tr><tr><td>小血管烧灼</td><td>精细科学工具</td><td> 18000-00 </td><td></td></tr><tr><td>明胶海绵</td><td>法玛西亚，辉瑞公司</td><td>调音台轧机MM400 </td><td></td></tr><tr><td>紧凑型脊髓 夹具</td><td> Narishige </td><td> STS - 一个</td><td></td></tr><tr><td>头部控股适配器</td><td> Narishige </td><td> MA - 6N </td><td></td></tr><tr><td> Gelseal </td><td> Amersham公司 生物科学公司</td><td> 80-6421-43 </td><td></td></tr><tr><td>乳酸钠林格</td><td>百特医疗用品&lt;/ TD&gt; <td> 2B8609 </td><td></td></tr><tr><td> Buprenex </td><td> Reckit洁 制药公司</td><td> NDC编号：12496 - 6757-1 </td><td>丁丙诺啡 注射</td></tr><tr><td> Baytril </td><td>拜耳</td><td> NADA 140-913 </td><td>恩诺沙星， 抗菌注射 2.27％（20毫升） </td></tr><tr><td>加热垫 - 大</td><td>精细科学工具</td><td> 21060-10 </td><td></td></tr></tbody></table>

in vivo imaging of the mouse spinal cord using two-photon microscopy

在体内成像已遗传工程小鼠表达荧光蛋白在特定的细胞类型 2-3显著扩大我们的生理和病理过程知识在众多的组织在 体内 4-7使用双光子显微镜 1 。在中枢神经系统（CNS）的研究，已被广泛的应用在体内成像在大脑中，这已经产生了众多新颖的细胞，如神经元，星形胶质细胞，小胶质细胞，在行为往往是意外的结果生理或病理条件下 8-17 。然而，大多是复杂的技术问题，有限的生活小鼠脊髓的研究在体内成像的实施。特别是，对脊髓的解剖毗邻的肺部和心脏产生显著运动伪影，使成像的生活脊髓一项艰巨的任务。 </P&gt; 我们开发出一种新的方法，克服了稳定脊柱，减少呼吸道引起的运动，从而促进利用双光子显微镜的图像在体内小鼠脊髓的脊髓造影固有的局限性。这是通过深麻醉方法相结合的一个定制的脊柱稳定装置，导致呼吸道引起的运动的一个显着减少。这个视频协议显示了如何公开一个可以维持稳定的生理条件下过长时间保持组织损伤和出血最低生活脊髓的小面积。具有代表性的原始图像在高分辨率收购体内详细的小胶质细胞和血管之间的密切关系。一个timelapse序列显示了在生活小鼠脊髓小胶质细胞过程的动态行为。此外，一个相同的Z -帧的连续扫描显示S的出色的稳定性，这种方法可以实现图像和/或timelapse电影，​​不需要图像对齐收购后产生的栈。最后，我们表明这种方法可用于如何重新审视和重新映像在稍后的时间点脊髓同一地区，从而在体内持续的生理或病理过程的纵向研究。

Watch this Scientific Journal Video about In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy at JoVE.com

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

微创的协议，以稳定脊柱鼠标和执行重复<em&gt;在体内</em&gt;脊髓成像，利用双光子显微镜描述。这种方法结合了脊柱的稳定装置和麻醉方案，以尽量减少引起的呼吸运动和生产没有对齐或其他处理后的原始图像数据，需要。

在体内脊髓的鼠标线使用双光子显微镜成像

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

Research

JoVE Journal

Neuroscience

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

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科研

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同时双光子<em&gt;在体内</em&gt;突触输入的影像和突触后目标中的鼠标Retrosplenial的Cortex

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