Name: single cell transfection in chick embryos
Uploaded: 2010-09-25T17:00:00
Description: Watch this Scientific Journal Video about Single Cell Transfection in Chick Embryos at JoVE.com

我们感谢施洛米Krispin神经管​​形象。特别是，我们承认尤瓦肉桂启动GFP转染技术。这项工作得到了来自以色列科学基金会，协会法国反对LES肌病，德意志Forschungsgemeinshaft，合作研究中心488的赠款为CK

1。准备的微量移液器我们Narishige PP - 83标准的吸管拉马拉我们的吸液管与热定型为13.5。精确的设置必须校准，目的是在枪头直径8至10微米。我们使用长丝含有硼硅玻璃管，外径1.0毫米，内径0.75-0.78毫米。移液器因此存储在一个塑料菜与菜的边缘接触保护提示。 2。 DNA的制备利益的质粒转染到大肠杆菌DH5α中使用标准程序的...

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	<li>Ben-Yair, R., Kalcheim, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+analysis+of+the+avian+dermomyotome+sheet+reveals+the+existence+of+single+cells+with+both+dermal+and+muscle+progenitor+fates.">Lineage analysis of the avian dermomyotome sheet reveals the existence of single cells with both dermal and muscle progenitor fates.</a> Development. 132, 689-701 (2005).</li><li>Ben-Yair, R., Kalcheim, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+and+bone+morphogenetic+protein+differentially+act+on+dermomyotome+cells+to+generate+endothelium,+smooth,+and+striated+muscle.">Notch and bone morphogenetic protein differentially act on dermomyotome cells to generate endothelium, smooth, and striated muscle.</a> J Cell Biol. 180, 607-618 (2008).</li><li>Cinnamon, Y., Kahane, N., Kalcheim, C. <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+early+development+of+specific+hypaxial+muscles+from+the+ventrolateral+myotome.+.">Characterization of the early development of specific hypaxial muscles from the ventrolateral myotome. .</a> Development. 126, 4305-4315 (1999).</li><li>Stark, D. A., Kulesa, 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=Photoactivatable+green+fluorescent+protein+as+a+single-cell+marker+in+living+embryos.">Photoactivatable green fluorescent protein as a single-cell marker in living embryos.</a> Dev Dyn. 233, 983-992 (2005).</li><li>Haas, K., Jensen, K., Sin, W. C., Foa, L., Cline, H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+electroporation+in+Xenopus+tadpoles+in+vivo--from+single+cells+to+the+entire+brain.">Targeted electroporation in Xenopus tadpoles in vivo--from single cells to the entire brain.</a> Differentiation. 70, 148-154 (2002).</li><li>Haas, K., Sin, W. C., Javaherian, A., Li, Z., Cline, H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+electroporation+for+gene+transfer+in+vivo.">Single-cell electroporation for gene transfer in vivo.</a> Neuron. 29, 583-591 (2001).</li><li>Hewapathirane, D. S., Haas, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+cell+electroporation+in+vivo+within+the+intact+developing+brain.">Single cell electroporation in vivo within the intact developing brain.</a> J Vis Exp. , (2008).</li><li>Nagai, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+variant+of+yellow+fluorescent+protein+with+fast+and+efficient+maturation+for+cell-biological+applications.">A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications.</a> Nat Biotechnol. 20, 87-90 (2002).</li><li>Sato, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+integration+and+conditional+expression+of+electroporated+transgenes+in+chicken+embryos.">Stable integration and conditional expression of electroporated transgenes in chicken embryos.</a> Dev Biol. 305, 616-624 (2007).</li></ol>

上面介绍的方法是修改为先前所描述的技术，提供亲脂性染料的小细胞亚群或3个单祖。它有三个以上的标准电技术的主要优势。首先，它提供了一个标签在靶组织中的信心离散网站的手段，让远远大于电空间分辨率（例如图1B，C）。其次，它允许单一细胞的标签，从而使跟踪的血统，另有 1 2（图1C，D） ​​的正常条件下在体内。三，基因表达怀念在单细胞水平宗族隔离，...

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single cell transfection in chick embryos

发育生物学的一个中心主题是多样化的谱系和澄清的分子机制。这需要一个正常和实验条件下的单细胞的命运的透彻分析。为此，目标单祖细胞的转染方法，是一个先决条件。我们在这里介绍一个简单的方法转染在OVO鸡组织中单个细胞，让可靠的谱系跟踪以及遗传操作技术。体节或神经管内，是针对特定的组织域的DNA直接注入利益的上皮细胞，导致零星的单细胞转染。记者，无性系种群可能因此被追踪长达三天，并与对照相比，无性系种群的基因操纵行为可以。此方法利用鸡胚无障碍的优势，以及与新兴的基因操纵的工具。我们比较，并讨论其优点被广泛使用的电击方法，以及可能的应用，还建议使用额外的体内模型。我们提倡使用这种方法的重要补充和补充现有的谱系追踪和基因干扰工具。

Watch this Scientific Journal Video about Single Cell Transfection in Chick Embryos at JoVE.com

Single Cell Transfection in Chick Embryos

使用精尖微量注入鸡体节或神经管的子域的质粒DNA。质粒的浓度调整，以产生单一的转染细胞。然后，我们允许的细胞发育成无性系种群。

单细胞转染鸡胚

A central theme in developmental biology is the diversification of lineages and the elucidation of underlying molecular mechanisms. This entails a ...

Research

JoVE Journal

Biology

Dissection of 6.5 dpc Mouse Embryos

6.5 DPC小鼠胚胎剖析

Analysis of gene expression patterns during early stages of mammalian embryonic development can provide important clues about gene function, cell-cell ...

科研

生物学

In Ovo Electroporations of HH Stage 10 Chicken Embryos

在OVO的HH阶段10个鸡胚Electroporations

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology.  With the advent of ...

Method for Culture of Early Chick Embryos ex vivo (New Culture)

Method for Culture of Early Chick Embryos ex vivo New Culture

文化早期鸡胚体外（新文化）的方法

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Double Whole Mount in situ Hybridization of Early Chick Embryos

双整装原位杂交技术在早期鸡胚

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

电子显微镜对胚胎的制备果蝇胚胎心管

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

Single-cell Suction Recordings from Mouse Cone Photoreceptors

单从小鼠锥感光细胞吸录音

Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment ...

Two-Photon-Based Photoactivation in Live Zebrafish Embryos

在现场的斑马鱼胚胎的基于双光子Photoactivati​​on

Photoactivation of target compounds in a living organism has proven a valuable approach to investigate various biological processes such as embryonic ...

Imaging Cell Shape Change in Living Drosophila Embryos

Imaging Cell Shape Change in Living Drosophila Embryos

影像生活细胞的形状变化果蝇胚胎

The developing Drosophila melanogaster embryo undergoes a number of cell shape changes that are highly amenable to live confocal imaging.  Cell shape ...

Single Cell Fate Mapping in Zebrafish

单细胞在斑马鱼的命运映射

The ability to differentially label single cells has important implications in developmental biology. For instance, determining how hematopoietic, ...

Facial Transplants in Xenopus laevis Embryos

Facial Transplants in Xenopus laevis Embryos

在面部移植<em&gt;非洲爪蟾</em&gt;胚胎

Craniofacial birth defects occur in 1 out of every 700 live births, but etiology is rarely known due to limited understanding of craniofacial development. ...