Name: preparation of drosophila s2 cells for light microscopy
Uploaded: 2010-06-03T17:00:00
Description: Watch this Scientific Journal Video about Preparation of Drosophila S2 cells for Light Microscopy at JoVE.com

这项工作是支持部分由美国国家癌症研究所P30 CA23074，美国癌症协会的机构研究资助74-001-31，和大学。亚利桑那胃肠孢子（NCI / NIH的CA9506O）。

1。 S2细胞显微镜前的准备工作施耐德S2细胞均来自俄勒冈ř 果蝇的胰酶消化后期胚胎。施耐德公司的原始文化类型的细胞的混合物，但变得更均匀继续通过1。他们已被描述为类似血细胞的基因表达巨噬细胞样（他们是吞噬）。 S2细胞可从ATCC，DGRC，或Invitrogen公司。 A.生长介质的选择多家媒体已用于文化S2细胞。媒体没有?...

<ol>
	<li>Schneider, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+lines+derived+from+late+embryonic+stages+of+Drosophila+melanogaster.">Cell lines derived from late embryonic stages of Drosophila melanogaster.</a> J Embryol Exp Morphol. 27, 353-365 (1972).</li><li>Rogers, S. L., Rogers, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culture+of+Drosophila+S2+cells+and+their+use+for+RNAi-mediated+loss-of-function+studies+and+immunofluorescence+microscopy.">Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy.</a> Nat Protoc. 3, 606-611 (2008).</li><li>Rogers, S. L., Rogers, G. C., Sharp, D. J., Vale, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+EB1+is+important+for+proper+assembly,+dynamics,+and+positioning+of+the+mitotic+spindle.">Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle.</a> J Cell Biol. 158, 873-884 (2002).</li><li>Goshima, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+required+for+mitotic+spindle+assembly+in+Drosophila+S2+cells.">Genes required for mitotic spindle assembly in Drosophila S2 cells.</a> Science. 316, 417-421 (2007).</li></ol>

对于细胞生物学领域中，一个理想的系统将保持成本低廉，易于操纵，并适合各种技巧。果蝇 S2细胞中满足这些要求，并让他们迅速成为越来越多的细胞的制度选择生物实验室。 我们提出了一个简要概述的方法，准备S2细胞显微镜。 S2细胞中的一个显着优点是他们成长，在正常的气氛和室温，因此，它们可以长时间没有任何特殊的维护要求的显微镜左。即使他们一般都...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>S2 cells</td>
<td>&nbsp;</td>
<td>ATCC</td>
<td>CRL-1963</td>
<td><a href="http://www.atcc.org/">http://www.atcc.org/</a></td>
</tr>
<tr>
<td>S2 cells</td>
<td>&nbsp;</td>
<td>DGRC</td>
<td>stock number 6</td>
<td><a href="https://dgrc.cgb.indiana.edu/">https://dgrc.cgb.indiana.edu/</a></td>
</tr>
<tr>
<td>S2 cells</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>R690-07</td>
<td>&nbsp;</td>
<tr>
<td>Sf900 II</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>10902-096</td>
<td>serum-free medium</td>
</tr>
<tr>
<td>HyClone SFX-Insect</td>
<td>&nbsp;</td>
<td>Thermo Scientific</td>
<td>SH30278</td>
<td>serum-free medium</td>
</tr>
<tr>
<td>Insect-Xpress</td>
<td>&nbsp;</td>
<td>BioWhittaker</td>
<td>12-730</td>
<td>serum-free medium</td>
</tr>
<tr>
<td>Schneider's S2 medium</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>11720-034</td>
<td>Add 10% (final) heat-inactivated FBS to make complete medium.</td>
</tr>
<tr>
<td>Fetal bovine serum</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>10438026</td>
<td>heat inactivated</td>
</tr>
<tr>
<td>100x antibiotic solution</td>
<td>&nbsp;</td>
<td>MP Biomedicals</td>
<td>91674049</td>
<td>contains penicillin (10000 U/mL), streptomycin (10000 &mu;g/mL), and amphotericin B (25 &mu;g/mL)</td>
</tr>
<tr>
<td>Concanavalin A</td>
<td>&nbsp;</td>
<td>MP Biomedicals</td>
<td>195283</td>
<td>&nbsp;</td>
<tr>
<td>Formaldehyde</td>
<td>&nbsp;</td>
<td>Electron Microscopy Sciences</td>
<td>15714</td>
<td>32% solution</td>
</tr>
<tr>
<td>Methanol</td>
<td>&nbsp;</td>
<td>Mallinckrodt Baker</td>
<td>9049</td>
<td>anhydrous, ACS grade</td>
</tr>
<tr>
<td>Molecular sieves</td>
<td>&nbsp;</td>
<td>Acros Organics</td>
<td>19724</td>
<td>type 3A</td>
</tr>		</tbody>
		</table>
		</div>

preparation of drosophila s2 cells for light microscopy

理想的实验系统，将便宜和易于维护，适合多种技术，将通过大量的文献和基因组序列数据库支持。培养的果蝇 S2细胞中，脱离20-24 小时 1岁胚胎的产品，具备所有这些特性。因此，S2细胞的细胞过程，包括发现的基因编码过程中的分子成分或利益机制的分析非常非常适合。其效用是最负责任的S2细胞的特点是与它们保持缓和，以其精湛的双链（DS）介导的RNA干扰（RNAi）的敏感性，以及居住或固定的可追踪性荧光显微镜细胞。 S2细胞可以生长在各种媒体，包括一些便宜，市售的无血清培养基，充分定义，2 。此外，他们成长优化，并迅速在21-24 ° C，可在各种容器中培养。哺乳动物细胞不同，S2细胞并不需要监管的氛围，而是与正常空气，甚至可以在保持密封烧瓶。 在S2细胞中的RNAi缓解的补充，是能够很容易分析实验诱导的表型相或固定或活细胞的荧光显微镜。文化作为一个单一的单层S2细胞的生长，但不显示接触抑制。相反，细胞往往生长在茂密的文化殖民地。在密度低，S2文化生长在玻璃或组织培养处理的塑料圆而松散连接。然而，S2细胞的细胞学检查可大大提高，诱使他们拉平简要地培养他们广泛的外源凝集素，刀豆蛋白A（ConA）3表面涂。 S2细胞中也可以稳定转染与荧光标记的标记，标签结构或细胞器在活细胞或固定细胞的兴趣。因此，细胞的微观分析通常的情况是这样的：第一，S2细胞（可以拥有转基因表达标签标记）通过RNAi治疗，以消除靶蛋白（S）。 RNA干扰治疗时间可以调整，以便在蛋白质的差异转动力学和，以减少细胞的创伤/死亡，如果目标蛋白是很重要的生存能力。接下来，处理后的细胞被转移到一盘菜含有盖玻片预涂与ConA诱导细胞扩散，紧紧坚持以玻璃。最后，细胞成像与研究者的显微镜模式的选择。 S2细胞，为需要扩展可视化活细胞的研究，特别是良好的，因为这些细胞停留在室温和常压下的健康。

Watch this Scientific Journal Video about Preparation of Drosophila S2 cells for Light Microscopy at JoVE.com

Preparation of Drosophila S2 cells for Light Microscopy

果蝇施耐德（S2）细胞基因的发现和功能分析系统越来越受欢迎。我们的目标是描述这样一个日益重要的实验系统，使S2细胞的显微技术。

制备果蝇 S2细胞，光学显微镜

The ideal experimental system would be cheap and easy to maintain, amenable to a variety of techniques, and would be supported by an extensive literature ...

Research

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Preparation of Drosophila S2 cells for Light Microscopy

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细胞器运输的培养<em&gt;果蝇</em&gt;细胞：S2细胞系和原代神经元。

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