Name: isolation of drosophila melanogaster testes
Uploaded: 2011-05-13T13:00:00
Duration: 3 min 13 s
Description: Watch this Scientific Journal Video about Isolation of Drosophila melanogaster Testes at JoVE.com

作者感谢Vasia Vagin这种技术引入到我们的实验室。

<ol><li>一个黑色背景的玻璃显微镜玻片上削减20＃白皮书同样大小的显微镜幻灯片。一种颜色的幻灯片尺寸纸张上的黑色方使用永久性标记（例如，Sharpie）。白胶（Elmer的）应用的黑边，坚持到幻灯片的文件。可重复使用的幻灯片。 ¾“标签的磁带（Fisherbrand），安全的显微镜载玻片，玻璃边，到CO 2飞垫修复到位。 </li><li>收集0-3日龄雄蝇。 </li><li>准备一个冰桶，磷酸盐缓冲液（PBS），解剖针。 </li><li>麻醉50-100使用的CO 2流的苍蝇和瓶或小瓶转移到附近的显微镜幻灯片飞垫。 </li><li>移液器〜50μL冰冷的PBS在显微镜幻灯片中下降。 </li><li>轻轻拿起一个男飞的解剖针DIP解剖针针的地方飞入的PBS下降到PBS，温柔的触摸腹部（即近的翅膀，）。 </li><li>飞中间放置一针，胸部和腹部之间，倒推坚决 。广场之间的第六届（A6）和腹节第七届（A7）（从底部的第二个和第三个条纹）其他针，腹部拉离胸部。 </li><li>睾丸除了其他器官，尤其是肠道挑逗。 ，尤其是在白色的突变体，睾丸和肠道类似于。主要区别是，睾丸总是成对出现;每个睾丸有螺旋纹，白色的比肠道。 </li><li>聚丙烯离心管中，在冰冷的PBS商店的睾丸。 </li></ol>代表性的成果： <img src="/files/ftp_upload/2641/2641fig1.jpg" alt="图1" /> 图1。男性生殖系统的背面观，从flybase.org/reports/FBim0000074.html。 Flybase归属：“米勒后，米勒（1950年）页508图38一个内部解剖和组织学的果蝇成虫在M. Demerec，编辑，果蝇的生物学，第6章，420页 - 534。冷泉港实验室出版社，传真版，1994年由冷泉港实验室出版社1994年版权使用一种来自冷泉港实验室（CSHL），许可按。 <a href="http://www.cshlpress.com">http://www.cshlpress.com</a> 。“

<ol>
	<li>Singh, S. R., Hou, S. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistological+Techniques+for+Studying+the+Drosophila+Male+Germline+Stem+Cell.">Immunohistological Techniques for Studying the Drosophila Male Germline Stem Cell.</a> Methods Mol Biol. 450, 45-59 (2008).</li></ol>

我们提供了协议隔离成年雄性果蝇睾丸。由于技术要求协调人的两只手，技能只有与实践。

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<td>Dissecting needle</td>
<td>Tool</td>
<td>Fisher Scientific</td>
<td>S17346</td>
<td>&nbsp;</td>
<tr>
<td>Micro Slides, Plain </td>
<td>Tool</td>
<td>Fisher Scientific</td>
<td>12-553</td>
<td>&nbsp;</td>
<tr>
<td>Microscope Light</td>
<td>Tool</td>
<td>Schott-Fostec</td>
<td>KO-1500</td>
<td>&nbsp;</td>
<tr>
<td>CO2 pad</td>
<td>Tool</td>
<td>Genesee Scientific</td>
<td>59-172</td>
<td>&nbsp;</td>
<tr>
<td>Benchtop Flowbuddy Complete System</td>
<td>Tool</td>
<td>Genesee Scientific</td>
<td>59-122WCU</td>
<td>&nbsp;</td>
<tr>
<td>Bubble Humidifier</td>
<td>Tool</td>
<td>Salter Labs</td>
<td>7100</td>
<td>&nbsp;</td>
<tr>
<td>Ball filler needle</td>
<td>Tool</td>
<td>Genesee Scientific</td>
<td>54-119</td>
<td>&nbsp;</td>
<tr>
<td>&frac34;&#x201D; Label Tape</td>
<td>Supply</td>
<td>Fisher Scientific</td>
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isolation of drosophila melanogaster testes

果蝇睾丸干细胞维持和分化，减数分裂和SOMA的生殖细胞相互作用的研究提供一个重要的模式。隔离成年男性的睾丸通常从蛹的情况下羽化后0-3天。野生型果蝇的睾丸很容易区别于其他组织的，因为他们是黄色的，但白突变的果蝇睾丸，一个共同的遗传背景的实验室实验飞肠道的形状和颜色相似。表演夹层玻璃显微镜玻片上，一个黑色的背景，使得相当容易识别的睾丸。睾丸被删除的苍蝇，用解剖针。使用睾丸清扫钳的协议相比，我们的方法是远远更快，允许一个良好的实践个人剖析每小时从200-300野生型果蝇睾丸，产生400-600睾丸。 白蝇，或从突变体，减少睾丸大小的睾丸更难解剖，通常每小时产量200-400睾丸。

Watch this Scientific Journal Video about Isolation of Drosophila melanogaster Testes at JoVE.com

Isolation of Drosophila melanogaster Testes

果蝇睾丸可迅速和有效地从成年男性使用解剖针分离。通过实践，人们可以很容易地在一至两天隔离睾丸足够的抗体或酶为基础的分析的高通量测序或更传统的分子生物学方法分析的DNA或RNA或蛋白质的量。

分离果蝇睾丸

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline ...

Research

JoVE Journal

Biology

Isolation of Drosophila melanogaster Testes

Dissection of Larval CNS in Drosophila Melanogaster

在果蝇幼虫中枢神经系统的剖析

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

科研

生物学

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

高分辨率视频跟踪移动的，在成年果蝇。

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

实时成像果蝇胚胎血细胞迁移

Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we ...

Dissection of Oenocytes from Adult Drosophila melanogaster

Dissection of Oenocytes from Adult Drosophila melanogaster

从成人Oenocytes剖析果蝇</em

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

寿命测量<em&gt;果蝇</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes

Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes

精子发生的细胞学分析：Live和固定制剂<em&gt;果蝇</em&gt;睾丸

Drosophila melanogaster is a powerful model system that has been widely used to elucidate a variety of biological processes. For example, studies of both ...

Imaging Through the Pupal Case of Drosophila melanogaster

Imaging Through the Pupal Case of Drosophila melanogaster

通过影像学的蛹壳<em&gt;果蝇</em

The longstanding use of Drosophila as a model for cell and developmental biology has yielded an array of tools. Together, these techniques have enabled ...

Dissection and Immunostaining of Imaginal Discs from Drosophila melanogaster

Dissection and Immunostaining of Imaginal Discs from Drosophila melanogaster

解剖和免疫染色成虫盘的<em&gt;果蝇</em

A significant portion of post-embryonic development in the fruit fly, Drosophila melanogaster, takes place within a set of sac-like structures called ...

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

增强Northern印迹检测小RNA种类的<em&gt;果蝇</em

The last decades have witnessed the explosion of scientific interest around gene expression control mechanisms at the RNA level. This branch of molecular ...

Maintenance of a Drosophila melanogaster Population Cage

Maintenance of a Drosophila melanogaster Population Cage

的维持<em&gt;果蝇</em&gt;人口笼

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life ...