Name: live dissection of drosophila embryos: streamlined methods for screening mutant collections by antibody staining
Uploaded: 2009-12-29T15:00:00
Description: Watch this Scientific Journal Video about Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining at JoVE.com

我们感谢他们的贡献，这些协议的发展Nicki福克斯和阿洛伊西亚施密德。这项工作是支持由NIH RO1授予KZ​​，NS28182。

简介 果蝇胚胎胚胎发育的阶段，14和17之间，可以很容易地解剖产生“鱼片”的准备工作。在这些准备工作，中枢神经系统（CNS）运行中间，是由身体墙壁两侧。肠道中被删除。当抗体染色，鱼片允许更好的可视化的中枢神经系统和体壁结构（如电机轴突，肌肉，周边感觉神经元（PNS）的，气管）比整个安装的胚胎，因为没有组织之间的准备和干预盖玻片，因为鱼...

<ol>
	<li>Patel, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+neuronal+subsets+and+other+cell+types+in+whole-mount+Drosophila+embryos+and+larvae+using+antibody+probes.">Imaging neuronal subsets and other cell types in whole-mount Drosophila embryos and larvae using antibody probes.</a> Methods Cell Biol. 44, 445-487 (1994).</li><li>Desai, C. J., Gindhart, J. G., Goldstein, L. S., Zinn, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Receptor+tyrosine+phosphatases+are+required+for+motor+axon+guidance+in+the+Drosophila+embryo.">Receptor tyrosine phosphatases are required for motor axon guidance in the Drosophila embryo.</a> Cell. 84, 599-609 (1996).</li><li>Sun, Q., Schindelholz, B., Knirr, M., Schmid, A., Zinn, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complex+genetic+interactions+among+four+receptor+tyrosine+phosphatases+regulate+axon+guidance+in+Drosophila.">Complex genetic interactions among four receptor tyrosine phosphatases regulate axon guidance in Drosophila.</a> Molecular and cellular neurosciences. 17, 274-291 (2001).</li><li>Schmid, A., Schindelholz, B., Zinn, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial+RNAi:+a+method+for+evaluating+the+functions+of+gene+families+in+Drosophila.">Combinatorial RNAi: a method for evaluating the functions of gene families in Drosophila.</a> Trends in neurosciences. 25, 71-74 (2002).</li><li>Fox, A. N., Zinn, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+heparan+sulfate+proteoglycan+syndecan+is+an+in+vivo+ligand+for+the+Drosophila+LAR+receptor+tyrosine+phosphatase.">The heparan sulfate proteoglycan syndecan is an in vivo ligand for the Drosophila LAR receptor tyrosine phosphatase.</a> Curr Biol. 15, 1701-1711 (2005).</li></ol>

我们希望这个视频演示，显示了我们的活胚胎解剖和任何人有一些在果蝇的遗传学和胚胎学的经验，他们现在可以很容易执行的足够详细的染色方法。当然，将仍然需要相当的实践，主要用于解剖自己。根据我们的经验，每个人学习生活的清扫方法，将创建一个微妙的不同剥离协议，允许他/她最快速的生成高质量的摆着解剖。这个过程需要全面发展的个月。然而，大多数人经过几个星期的...

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<td>Borosilicate Tubing With Filament </td>
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live dissection of drosophila embryos: streamlined methods for screening mutant collections by antibody staining

果蝇胚胎胚胎发育的阶段，14和17之间，可以很容易地解剖产生“鱼片”的准备工作。在这些准备工作，中枢神经系统运行的中间，是由身体墙壁两侧。许多不同表型已使用这种制剂的研究。鱼片在大多数情况下，所产生的抗体染色的固定安装整个胚胎的夹层。这些“固定的解剖”，但有一些缺点。他们耗费时间来执行，也很难在保持了GFP平衡器染色体突变的股票进行排序的突变体（GFP阴性）胚胎。自2002年以来，本集团一直在进行缺乏症和异位表达屏幕确定为孤儿受体的配体。为了做到这一点，我们开发的精简协议活胚胎剥离和抗体染色包含数百个平衡线路的集合。我们得出的结论，这是相当多的效率比固定清扫现场清扫研究股票的大集合的表型。使用这里描述的协议，单一的训练有素的个体可以通过屏幕10号线每天的表型，从每行的复合显微镜下检查4-7突变胚胎。这使得查明赋予微妙，低外显的表型的基因突变，在高放大倍率拿下了一个40倍的水浸泡镜片，因为每行最多70 hemisegments。

Watch this Scientific Journal Video about Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining at JoVE.com

Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining

我们描述了一个用于生成特定基因型的果蝇胚胎的“鱼片”筹备工作的精简的协议。此协议允许多种遗传筛选的高效执行。它也可以让优秀的中后期的胚胎结构的可视化。

现场剖析果蝇胚胎抗体染色筛选突变体集合的简化方法

Drosophila embryos between stages 14 and 17 of embryonic development can be readily dissected to generate "fillet" preparations.  In these preparations, ...

Research

JoVE Journal

Biology

Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining

Dissection of Drosophila Ovaries

剖析果蝇卵巢

科研

生物学

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 ...

Method for Whole Mount Antibody Staining in Chick

整装在鸡的抗体染色方法

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

Drosophila Larval NMJ Dissection

果蝇幼虫NMJ夹层

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use ...

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

编制果蝇实时成像的胚胎使用悬滴协议

Green fluorescent protein (GFP)-based timelapse live-imaging is a powerful technique for studying the genetic regulation of dynamic processes such as ...

Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling

果蝇聚乙烯南瓜抗体标记染色体的制备

Drosophila has long been a favorite model system for studying the relationship between chromatin structure and gene regulation due to the cytological ...

In-vivo Centrifugation of Drosophila Embryos

In-vivo Centrifugation of Drosophila Embryos

果蝇胚胎的体内离心

A major strategy for purifying and isolating different types of intracellular organelles is to separate them from each other based on differences in ...

Upright Imaging of Drosophila Embryos

Upright Imaging of Drosophila Embryos

直立成像果蝇胚胎

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current ...

RNAi Interference by dsRNA Injection into Drosophila Embryos

RNAi Interference by dsRNA Injection into Drosophila Embryos

RNAi的干扰双链RNA注射到果蝇胚胎

Genetic screening is one of the most powerful methods available for gaining insights into complex biological process 1. Over the years many improvements ...

Dissection and Staining of Drosophila Larval Ovaries

Dissection and Staining of Drosophila Larval Ovaries

夹层和染色果蝇幼虫卵巢

Many organs depend on stem cells for their development during embryogenesis and for maintenance or repair during adult life. Understanding how stem cells ...