Name: mouse eye enucleation for remote high-throughput phenotyping
Uploaded: 2011-11-19T14:00:00
Description: Watch this Scientific Journal Video about Mouse Eye Enucleation for Remote High-throughput Phenotyping at JoVE.com

研究以预防失明，主任朱尔斯·斯坦眼科研究所，加州大学洛杉矶分校Bartly J. Mondino MD和拉米罗·拉米雷斯 - 索利斯，杰奎白，和珍妮Estabel的，威康信托基金会桑格研究所基因组校园。本研究符合ARVO声明眼科和视觉研究中使用动物。

1。在不固定的标本钝性分离：老鼠的眼睛剜除术<ol><li>拉开眼睑改善及接触后的地球（眼球）表面。 </li><li>将弯曲敷料镊子背后（下）的地球轨道（眼圈）。马哈詹Sharptip敷料镊子尖这一步更容易（见材料和​​试剂表）是一个自定义的仪器。 </li><li>关闭的镊子和把握的轨道结缔组织和视神经背后的全球，同时小心避免挤压全球。 </li><li>轻轻向上拉镊子和勇敢的眼球从眼眶。白线样组织是视?...

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	<li>Song, B. J., Tsang, S. H., Lin, C. -. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+models+of+retinal+degeneration+and+targets+for+gene+therapy.">Genetic models of retinal degeneration and targets for gene therapy.</a> Gene Ther. Mol. Biol. 11, 229-262 (2007).</li><li>Mahajan, V. B., Mondino, B. J., Tsang, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-throughput+Mouse+Eye+Phenomics+System.">A high-throughput Mouse Eye Phenomics System.</a> Cold Spring Harbor Laboratories, Mouse Genetics Meeting. , (2010).</li><li>Smith, R. S., John, S. W. M., Nishina, P. M., Sundberg, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In Research Methods For Mutant Mice. , (2002).</li><li>Chang, B., Hawes, N. L., Hurd, R. E., Davisson, M. T., Nusinowitz, S., Heckenlively, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+degeneration+mutants+in+the+mouse.">Retinal degeneration mutants in the mouse.</a> Vision Res. 42, 517-525 (2002).</li><li>Anderson, M. G., Smith, R. S., Hawes, N. L., Zabaleta, A., Chang, B., Wiggs, J. L., John, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+in+genes+encoding+melanosomal+proteins+cause+pigmentary+glaucoma+in+DBA/2J+mice.">Mutations in genes encoding melanosomal proteins cause pigmentary glaucoma in DBA/2J mice.</a> Nat. Genet. 30, 81-85 (2002).</li><li>Hawes, N. L., Smith, R. S., Chang, B., Davisson, M., Heckenlively, J. R., John, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+fundus+photography+and+angiography:+a+catalogue+of+normal+and+mutant+phenotypes.">Mouse fundus photography and angiography: a catalogue of normal and mutant phenotypes.</a> Mol. Vis. 5, 22-22 (1999).</li><li>Won, J., Shi, L. Y., Hicks, W., Wang, J., Hurd, R., Naggert, J. K., Chang, B., Nishina, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+resources+for+vision+research.">Mouse model resources for vision research.</a> J. Ophthalmol. 2011, 391384-391384 (2011).</li><li>Pinto, L. H., Vitaterna, M. H., Siepka, S. M., Shimomura, K., Lumayag, S., Baker, M., Fenner, D., Mullins, R. F., Sheffield, V. C., Stone, E. M., Heffron, E., Takahashi, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Results+from+screening+over+9000+mutation-bearing+mice+for+defects+in+the+electroretinogram+and+appearance+of+the+fundus.">Results from screening over 9000 mutation-bearing mice for defects in the electroretinogram and appearance of the fundus.</a> Vision. Res. 44, 3335-3345 (2004).</li><li>Heckenlively, J. R., Winston, J. V., Roderick, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Screening+for+mouse+retinal+degenerations.+I.+Correlation+of+indirect+ophthalmoscopy,+electroretinograms,+and+histology.">Screening for mouse retinal degenerations. I. Correlation of indirect ophthalmoscopy, electroretinograms, and histology.</a> Doc. Ophthalmol. 71, 229-239 (1989).</li><li>Tsang, S. H., Gouras, P., Yamashita, C. K., Kjeldbye, H., Fisher, J., Farber, D. B., Goff, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+degeneration+in+mice+lacking+the+gamma+subunit+of+the+rod+cGMP+phosphodiesterase.">Retinal degeneration in mice lacking the gamma subunit of the rod cGMP phosphodiesterase.</a> Science. 272, 1026-1029 (1996).</li><li>Tsang, S. H., Woodruff, M. L., Jun, L., Mahajan, V., Yamashita, C. K., Pedersen, R., Lin, C. S., Goff, S. P., Rosenberg, T., Larsen, M., Farber, D. B., Nusinowitz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+mice+carrying+the+H258N+mutation+in+the+gene+encoding+the+beta-subunit+of+phosphodiesterase-6+(PDE6B)+provide+a+model+for+human+congenital+stationary+night+blindness.">Transgenic mice carrying the H258N mutation in the gene encoding the beta-subunit of phosphodiesterase-6 (PDE6B) provide a model for human congenital stationary night blindness.</a> Hum. Mutat. 28, 243-254 (2007).</li></ol>

大多数转基因小鼠中存在的不检查眼睛的实验室。我们的视频技术，说明了一个简单的，标准化的方法，用于远程手术技巧转移到优化组织收购，从实验室经验少用眼。此视频技术可以帮助克服在高通量的表型，这是由于非标准化组织收集和固定方法，防止有意义的比较表型和分子生物学研究的专业网站的使用数量有限的一个主要缺陷。任何新的远程实验室和技术人员，建立和保持质量控制，重?...

<table border="1"><tbody><tr><th>名称</th><th>公司</th><th>目录编号</th><th>评论</th></tr><tr><td>弧形修整镊子</td><td> STORZ眼药水</td><td> E1408 </td><td></td></tr><tr><td>马哈詹Sharptip敷料镊子</td><td> STORZ眼药水</td><td> E1406（SP7-64520 REF） </td><td></td></tr><tr><td>弯曲韦斯科特剪刀</td><td> STORZ眼药水</td><td> E3321 WH </td><td></td></tr><tr><td> 15°BD海狸显微叶片</td><td>贝克顿 - 迪金森</td><td> 374881 </td><td></td></tr><tr><td> 0.22精细Castroviejo的缝合钳</td><td> STORZ眼药水</td><td> E1805 </td><td></td></tr><tr><td> 0.12科利柏钳</td><td> STORZ眼药水</td><td>一百三十二分之二</td><td></td></tr><tr><td> 30号针头</td><td>贝克顿 - 迪金森</td><td> 305128 </td><td></td></tr><tr><td>生化危机梅勒</td><td>费舍尔</td><td> 03-523-4 </td><td></td></tr><tr><td>封口膜</td><td>费舍尔</td><td> 13-374-10 </td><td></td></tr><tr><td>玻璃闪烁瓶</td><td>惠顿</td><td> 4500413033 </td><td></td></tr><tr><td> PBS，pH值7.4 </td><td> Invitrogen公司</td><td> 70011-044 </td><td></td></tr><tr><td> 16％的多聚甲醛</td><td>电子显微镜科学</td><td> 15700 </td><td></td></tr><tr><td> 2.5％的多聚甲醛/ 2.5％戊二醛的0.1M磷酸钠缓冲液中</td><td>电子显微镜科学</td><td> 15700和16300 </td><td>混合实验室</td></tr><tr><td> 50％的戊二醛</td><td>电子显微镜科学</td><td> 16300 </td><td></td></tr><tr><td> 0.25％福尔瓦， </td><td>电子显微镜科学</td><td> 15810 </td><td></td></tr><tr><td>警察每插槽电网</td><td>电子显微镜科学</td><td> M2010-CR </td><td></td></tr><tr><td> 4％的四氧化锇</td><td>电子显微镜科学</td><td> 19140 </td><td></td></tr><tr><td>反SOD3抗体</td><td> Abcam公司</td><td> Ab21974 </td><td></td></tr><tr><td>山羊抗兔的Alexa Fluor 488 </td><td> Invitrogen公司</td><td> A11070 </td><td></td></tr><tr><td>斯珀尔的嵌入树脂</td><td>电子显微镜科学</td><td> 14300 </td><td></td></tr></tbody></table>

mouse eye enucleation for remote high-throughput phenotyping

老鼠的眼睛是一个重要的遗传模型的平移人类眼科疾病的研究。在转基因小鼠在人类的致盲性疾病，如黄斑变性，光感受器变性，白内障，青光眼，视网膜母细胞瘤，糖尿病性视网膜病变已概括。1-5大多数转基因和基因敲除小鼠的实验室研究已产生非眼科疾病，但器官系统之间的遗传保护，许多相同的基因也可能在眼发育和疾病发生中发挥作用。因此，这些老鼠的眼睛发现新的基因型与表型的相关性是一个重要的资源。由于这些老鼠分散在世界各地，它是难以获取，维护和​​表型以高效，成本效益的方式。因此，大多数高通量眼表型屏幕被限制在检查现场，眼科专业知识，需要几个地点的眼睛在活体小鼠6-9本实验室所开发的另一种方法是远程组织收购，可用于转基因小鼠的眼睛在或大或小规模的调查方法。基于视频的手术技术转移，组织固定和运输的标准化程序允许任何实验室收集整个眼睛突变的动物，并将它们发送的分子和形态学表型。在这个视频文章中，我们提出了远程表型分析技术enucleate和转让不固定，并灌注固定鼠标的眼睛。

Watch this Scientific Journal Video about Mouse Eye Enucleation for Remote High-throughput Phenotyping at JoVE.com

Mouse Eye Enucleation for Remote High-throughput Phenotyping

的解剖技术说明组织固定，进行表型的高通量筛选老鼠的眼睛剜除术。

老鼠的眼睛剜除术的远程高通量的表型

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