Name: chromosomics: detection of numerical and structural alterations in all 24 human chromosomes simultaneously using a novel octochrome fish assay
Uploaded: 2012-02-06T13:00:00
Description: Watch this Scientific Journal Video about Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay at JoVE.com

作者希望感谢博士Cliona研究麦克海尔批判性阅读和编辑的稿件。这项工作是由美国国家环境健康科学，国家卫生研究所授予R01ES017452张为L研究所。

1。样品幻灯片准备<ul><li> OctoChrome鱼，旨在探讨在人体细胞，包括中期利差的染色体变化，但不限于外周血细胞培养，干/祖细胞和细胞株。样品应准备后的收获中期3,11-13的标准程序：采用秋水仙胺治疗逮捕中期，低渗处理的细胞肿胀细胞，卡诺的固定液固定在悬挂在约0.5-1×10 6个细胞的细胞每毫升。 </li><li>清理的8平方米的幻灯片（中的OctoChrome的套件，目录＃：大马803 Cytocell有...

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	<li>Levsky, J. M., Singer, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+in+situ+hybridization:+past,+present+and+future.">Fluorescence in situ hybridization: past, present and future.</a> J. Cell. Sci. 116, 2833-2838 (2003).</li><li>Zhang, L., Eastmond, D. A., Smith, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+nature+of+chromosomal+aberrations+detected+in+humans+exposed+to+benzene.">The nature of chromosomal aberrations detected in humans exposed to benzene.</a> Crit. Rev. 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这种新颖的OctoChrome鱼法允许同时在一张幻灯片上的单一杂交研究所有人类染色体24。 8平方的染色体组合的安排，已设计，以方便识别中最常见的白血病和淋巴瘤的非随机的染色体重排。因此，该法可检测数值（非整倍体）以及结构（易位）染色体的改变同时进行。 在这个实验中最关键的一步是控制湿度在过夜杂交在水浴浮动商会。 FISH探针和杂交缓冲容易干燥，如果湿度低?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td><tr><tr><td> OctoChrome套件</td><td> cytocell有限公司，英国剑桥</td><td>大马803 </td><td></td></tr><tr><td>植物血凝素（PHA的） </td><td> Invitrogen公司</td><td> 10576-015 </td><td></td></tr><tr><td>秋水仙胺</td><td> Invitrogen公司</td><td> 15212-012 </td><td></td></tr><tr><td>卡诺的固定液</td><td></td><td></td><td>甲醇：冰醋酸= 3：1 </td></tr><tr><td>荧光公里croscope </td><td></td><td></td><td>配备过滤器，以查看德州红，FITC标记，香豆素光谱单独和的DAPI / FITC /得克萨斯州红三重过滤的DAPI，同时查看不同颜色</td></tr><tr><td> metafer软件</td><td> MetaSystems，Altlussheim，德国</td><td></td><td>方便找到所有中期和重新评估异常</td></tr></tbody></table>

chromosomics: detection of numerical and structural alterations in all 24 human chromosomes simultaneously using a novel octochrome fish assay

荧光原位杂交（FISH）是一种技术，使特定的DNA序列，中期或相间染色体检测细胞核1。该技术采用独特的序列杂交，整条染色体或特定的染色体区域的DNA探针，作为一个强大的辅助经典遗传学。例如，许多早期的研究报告中增加相关的白血病患者接触苯，苯中毒患者，接触苯的健康工人的染色体畸变的频繁检测，采用经典遗传学分析2。使用鱼，已观察到白血病特定染色体改变升高，显然健康的工人暴露于苯3-6，说明在苯致白血病细胞遗传学改变的关键角色。 一般来说，一条鱼检测只检查一个或几个整条染色体或每张幻灯片的特定基因，使多个杂交需要进行多张幻灯片，涵盖所有人类染色体。光谱核型分析（SKY）允许可视化的整个基因组的同时，但要求特殊的软件和设备的限制及其应用7。在这里，我们描述了一种新的鱼类试验，OctoChrome鱼，可用于Chromosomics的应用，这是我们定义为所有人类染色体24上的一个广泛的染色体非整倍体的研究，人类的研究，如幻灯片同步分析（CWAS） 8。作为Chromoprobe多功能系统由Cytocell上市的方法，基础是OctoChrome的的设备，被划分成8个广场，每个带有三种不同的整条染色体涂染探针（图1）。三个探头都直接标有不同颜色的荧光，绿色（FITC标记），红（得克萨斯红），蓝（香豆素）。染色体组合的安排上OctoChromE设备已设计，以方便识别中最常见的白血病和淋巴瘤中发现的非随机的染色体结构改建（易位），实例t（9; 22），T（15 17），T（8; 21 ），T（14 18）9。此外，在染色体上的数值变化（非整倍体）可以同时检测。相应的模板幻灯片，也分为8到中期利差是约束（图2），定位在OctoChrome设备的平方。在高温和潮湿室杂交探针和目标DNA变性，然后所有人类染色体24同时可以可视化。 OctoChrome FISH技术是一种很有前途的技术为白血病和淋巴瘤的临床诊断和所有染色体的非整倍体的检测。我们已申请在研究苯暴露中国工人8,10 CWAS这个新Chromosomic方法。

Watch this Scientific Journal Video about Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay at JoVE.com

Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay

一种新型的荧光<em&gt;原位</em&gt;杂交（FISH）的方法，同时检查数值和染色体结构的改变，尤其是特定的染色体易位的白血病和淋巴瘤相关的所有24人类染色体上的一​​个杂交的单设备，描述。

在所有人类染色体24 chromosomics：检测的数值模拟和结构上的改动同时使用一种新型OctoChrome鱼含量

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