Name: labeling and imaging cells in the zebrafish hindbrain
Uploaded: 2010-07-25T17:00:00
Description: Watch this Scientific Journal Video about Labeling and Imaging Cells in the Zebrafish Hindbrain at JoVE.com

这项工作是由美国国立卫生研究院授予授予R.布鲁斯特（1R01GM085290 - 01A1）支持。

1.Microinjection <ol><li>稀释质粒编码的膜有针对性的绿色荧光蛋白（mGFP，礼貌理查德哈兰德）至40毫微克/毫升的浓度。 DNA是由线性质粒（mGFP/PCS2，礼貌理查德哈兰德）纯化使用Qiagen公司马克西准备包准备。酚红（稀释后总体积的1 / 10）的解决方案是首选的可视化解决方案。 DNA溶液置于冰上。 </li><li>在立体显微镜下，校准的注射针（90毫米Narishinge科学的玻璃毛细管，猫编号NA - GD - 1），对准在毕业...

<ol>
	<li>N&#252;sslein-Volhard, C., Dahm, 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> Zebrafish Practical approach series. , (2002).</li><li>Hong, E., Brewster, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=N-cadherin+is+required+for+the+polarized+cell+behaviors+that+drive+neurulation+in+the+zebrafish.">N-cadherin is required for the polarized cell behaviors that drive neurulation in the zebrafish.</a> Development. 133, 3895-3905 (2006).</li><li>Andersen, E., Asuri, N., Clay, M., Halloran, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+Imaging+of+Cell+Motility+and+Actin+Cytoskeleton+of+Individual+Neurons+and+Neural+Crest+Cells+in+Zebrafish+Embryos.">Live Imaging of Cell Motility and Actin Cytoskeleton of Individual Neurons and Neural Crest Cells in Zebrafish Embryos.</a> J Vis Exp. , (2010).</li></ol>

总之，描述标签技术在这里允许在斑马鱼的胚胎形成过程的单细胞分析。此协议的主要重点是对成像标记在使用一个无处不在的启动子控制下的mGFP的神经管细胞的方法。对于这种瞬时表达检测读者的额外的应用程序应该是指一个由安德森等人最近的一篇文章 。3 。

A correction was made to <a href="http://www.jove.com/index/Details.stp?ID=1976">Labeling and Imaging Cells in the Zebrafish Hindbrain</a>. There was an error in the authors affiliations. The authors have been corrected to:
Pradeepa Jayachandran1, Elim Hong2, Rachel Brewster1 1Department of Biological Sciences, University of Maryland, Baltimore County 2Center for Neuroscience, Children's National Medical Center
instead of:
Pradeepa Jayachandran1, Elim Hong2, Rachel Brewster2 1Center for Neuroscience, Children's National Medical Center 2Department of Biological Sciences, University of Maryland Baltimore County

A correction to the author affiliations has been made for the article Labeling and Imaging Cells in the Zebrafish Hindbrain.

Erratum: Labeling and Imaging Cells in the Zebrafish Hindbrain

解决方案<ul><li>汉克的股票＃1：8.0 g氯化钠，氯化钾0.4克在100毫升DD H 2 O </li><li>汉克的联合＃2：0.358克钠2 HPO 4无水，0.60克KH 2 PO 4 100毫升DDH 2 O </li><li>汉克的股票＃4：0.72克氯化钙50毫升DDH 2 O 2 </li><li>汉克的股票5：1.23克硫酸镁 x7H 2 O在50毫升DD H 2 O </li><li> 1 + X磷酸盐缓冲液：0.8％氯化钠，0.02％氯化钾，0.02 M磷酸盐，pH值7.3 </li><li>封闭液：2％正常山羊血清，1％牛血清白蛋白（BSA），1％dimethysulfoxide（DMSO） </li></ul>

labeling and imaging cells in the zebrafish hindbrain

理解，塑造了早期脊椎动物胚胎的形态发生过程的关键是对高分辨率图像细胞的能力。在斑马鱼胚胎注射，镶嵌表达质粒DNA结果，使细胞的单细胞或小群的可视化<sup&gt; 1</sup&gt;。我们描述了如何注射质粒DNA编码一个无处不在的启动子控制下的膜有针对性的绿色荧光蛋白（mGFP）可用于神经胚细胞成像。这个协议的核心是节和实时高分辨率成像标记细胞的方法。该协议涉及mGFP的DNA注入到年轻的斑马鱼胚胎。胚胎，然后vibratome切片，抗体标记和共聚焦显微镜成像处理。另外，表达mGFP的活胚胎，可使用时间推移共聚焦显微镜成像。我们以前使用这种简单的方法来分析的驱动器在斑马鱼胚胎的后脑地区的神经管形成的细胞行为<sup&gt; 2</sup&gt;。允许前所未有的可视化细胞的形状和组织中的神经管，而这种方法使一个更好地了解在神经胚细胞动力学实时成像补充固定的准备。

Watch this Scientific Journal Video about Labeling and Imaging Cells in the Zebrafish Hindbrain at JoVE.com

Labeling and Imaging Cells in the Zebrafish Hindbrain

理解，塑造了早期胚胎的形态发生过程的关键是对高分辨率图像细胞的能力。我们在这里描述膜有针对性的绿色荧光蛋白标签在整个斑马鱼的胚胎细胞的单细胞或小群的技术。

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