Name: chromogenic in situ hybridization as a tool for hpv-related head and neck cancer diagnosis
Uploaded: 2019-06-14T13:00:00
Description: Watch this Scientific Journal Video about Chromogenic In Situ Hybridization as a Tool for HPV-Related Head and Neck Cancer Diagnosis at JoVE.com

作者感谢欧洲霍皮塔兰·乔治·蓬皮杜和内克尔病理学系(劳里安·尚博勒、埃洛迪·米歇尔和吉塞勒·勒格尔);PARCC组织学平台,欧洲霍皮塔·乔治·蓬皮杜(科林娜·莱萨弗雷);弗吉尼亚克拉克语言编辑;亚历山德拉·埃尔巴基扬为她的贡献。

该议定书遵循道德准则,并经道德委员会(第二次道德保护委员会,#2015-09-04)批准。
1. 材料的准备
<ol><li>
准备 1x 洗涤缓冲液
	<ol>
<li>通过在大型车童中加入 2.94 L 蒸馏水和一瓶(60 mL)洗涤缓冲液(50x)(参见材料表),制备 3 L 的 1x 洗涤缓冲液。混合好。 注:1x洗涤缓冲液可提前准备,并在室温下储存长达1个月。</li>
	</ol>
</li>
	<li>
反染色试剂的制备</strong...

<ol>
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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNA+chromogenic+in+situ+hybridization+assay+with+clinical+automated+platform+is+a+sensitive+method+in+detecting+high-risk+human+papillomavirus+in+squamous+cell+carcinoma.">RNA chromogenic in situ hybridization assay with clinical automated platform is a sensitive method in detecting high-risk human papillomavirus in squamous cell carcinoma.</a> Human Pathology. 63, 184-189 (2017).</li><li>Mirghani, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+of+HPV+driven+oropharyngeal+cancers:+Comparing+p16+based+algorithms+with+the+RNAscope+HPV-test.">Diagnosis of HPV driven oropharyngeal cancers: Comparing p16 based algorithms with the RNAscope HPV-test.</a> Oral Oncology. 62, 101-108 (2016).</li><li>Lewis, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+Papillomavirus+Testing+in+Head+and+Neck+Carcinomas:+Guideline+From+the+College+of+American+Pathologists.">Human Papillomavirus Testing in Head and Neck Carcinomas: Guideline From the College of American Pathologists.</a> Archives of Pathology &amp; Laboratory Medicine. 142 (5), 559-597 (2018).</li><li>Mills, A. M., Coppock, J. D., Willis, B. C., Stoler, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPV+E6/E7+mRNA+In+Situ+Hybridization+in+the+Diagnosis+of+Cervical+Low-grade+Squamous+Intraepithelial+Lesions+(LSIL).">HPV E6/E7 mRNA In Situ Hybridization in the Diagnosis of Cervical Low-grade Squamous Intraepithelial Lesions (LSIL).</a> The American Journal of Surgical Pathology. 42 (2), 192-200 (2018).</li><li>El-Naggar, A., Chan, J. K. C., Grandis, J. R., Takata, T., Slootweg, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> WHO Classification of Head and Neck Tumours. , (2017).</li><li>Ang, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+papillomavirus+and+survival+of+patients+with+oropharyngeal+cancer.">Human papillomavirus and survival of patients with oropharyngeal cancer.</a> The New England Journal of Medicine. 363 (1), 24-35 (2010).</li><li>Badoual, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PD-1-expressing+tumor-infiltrating+T+cells+are+a+favorable+prognostic+biomarker+in+HPV-associated+head+and+neck+cancer.">PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer.</a> Cancer Research. 73 (1), 128-138 (2013).</li><li>Outh-Gauer, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunotherapy+in+head+and+neck+cancers:+a+new+challenge+for+immunologists,+pathologists+and+clinicians.">Immunotherapy in head and neck cancers: a new challenge for immunologists, pathologists and clinicians.</a> Cancer Treatment Reviews. 65, 54-64 (2018).</li><li>Mirghani, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+of+HPV-driven+head+and+neck+cancer+with+a+single+test+in+routine+clinical+practice.">Diagnosis of HPV-driven head and neck cancer with a single test in routine clinical practice.</a> Modern Pathology. 28 (12), 1518-1527 (2015).</li><li>Bishop, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+Transcriptionally+Active+High-risk+HPV+in+Patients+With+Head+and+Neck+Squamous+Cell+Carcinoma+as+Visualized+by+a+Novel+E6/E7+mRNA+In+Situ+Hybridization+Method.">Detection of Transcriptionally Active High-risk HPV in Patients With Head and Neck Squamous Cell Carcinoma as Visualized by a Novel E6/E7 mRNA In Situ Hybridization Method.</a> The American Journal of Surgical Pathology. 36 (12), 1874-1882 (2012).</li><li>Hsieh, M. -. S., Lee, Y. -. H., Jin, Y. -. T., Huang, W. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strong+SOX10+expression+in+human+papillomavirus-related+multiphenotypic+sinonasal+carcinoma:+report+of+6+new+cases+validated+by+high-risk+human+papillomavirus+mRNA+in+situ+hybridization+test.">Strong SOX10 expression in human papillomavirus-related multiphenotypic sinonasal carcinoma: report of 6 new cases validated by high-risk human papillomavirus mRNA in situ hybridization test.</a> Human Pathology. 82, 264-272 (2018).</li><li>Shi, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+Prognostic+Value+of+HPV16+E6+mRNA+Compared+With+In+Situ+Hybridization+for+Human+Oropharyngeal+Squamous+Carcinoma.">Comparative Prognostic Value of HPV16 E6 mRNA Compared With In Situ Hybridization for Human Oropharyngeal Squamous Carcinoma.</a> Journal of Clinical Oncology. 27 (36), 6213-6221 (2009).</li><li>Kuo, K. -. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biomarkers+of+human+papillomavirus+infection+in+tonsillar+squamous+cell+carcinoma&#8212;molecular+basis+and+predicting+favorable+outcome.">The biomarkers of human papillomavirus infection in tonsillar squamous cell carcinoma&#8212;molecular basis and predicting favorable outcome.</a> Modern Pathology. 21 (4), 376-386 (2008).</li><li>Augustin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+efficacy+of+the+four+tests+(p16+immunochemistry,+PCR,+DNA+and+RNA+In+situ+Hybridization)+to+evaluate+a+Human+Papillomavirus+infection+in+head+and+neck+cancers:+a+cohort+of+348+French+squamous+cell+carcinomas.">Evaluation of the efficacy of the four tests (p16 immunochemistry, PCR, DNA and RNA In situ Hybridization) to evaluate a Human Papillomavirus infection in head and neck cancers: a cohort of 348 French squamous cell carcinomas.</a> Human Pathology. , (2018).</li><li>Jung, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+and+clinical+relevance+of+transcriptionally+active+human+papillomavirus+(HPV)+infection+in+oropharynx+squamous+cell+carcinoma.">Biological and clinical relevance of transcriptionally active human papillomavirus (HPV) infection in oropharynx squamous cell carcinoma.</a> International Journal of Cancer. 126 (8), 1882-1894 (2009).</li><li>Augustin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPV+RNA+CISH+score+identifies+two+prognostic+groups+in+a+p16+positive+oropharyngeal+squamous+cell+carcinoma+population.">HPV RNA CISH score identifies two prognostic groups in a p16 positive oropharyngeal squamous cell carcinoma population.</a> Modern Pathology. , (2018).</li><li>Evans, M. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPV+E6/E7+RNA+In+Situ+Hybridization+Signal+Patterns+as+Biomarkers+of+Three-Tier+Cervical+Intraepithelial+Neoplasia+Grade.">HPV E6/E7 RNA In Situ Hybridization Signal Patterns as Biomarkers of Three-Tier Cervical Intraepithelial Neoplasia Grade.</a> PLOS ONE. 9 (3), e91142 (2014).</li><li>Dreyer, J. H., Hauck, F., Oliveira-Silva, M., Barros, M. H. 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HPV RNA CISH 使用购买的试剂盒执行,是检测病毒记录组的强大工具,它表明 HPV 感染活跃。手动执行,协议的步骤整体上易于遵循,并且购买的工具包很方便。该技术允许一次染色19个组织学样本和一个控制幻灯片,测定持续约8小时。除非另有说明,否则不要让样品在步骤之间干燥,这一点至关重要。预处理条件可能需要根据操纵的组织进行调整。
HPV E6-E7 mRNA 表达信号以精确的空间分辨?...

HPV RNA CISH 是检测活动 HPV 感染的有力工具,在诸如口腔或子宫颈等不同部位的良性或恶性病变中可能证明至关重要。活动HPV感染的检测可能支持HPV引起的病变的诊断,从而影响其治疗和预后。
HPV是最常见的性传播感染,超过100种病毒基因型已被描述为1。从原理上讲,低风险基因型(如基因型6和11)已知可诱发生殖器疣、复发性呼吸道性手皮瘤病和其他良性病变,而高风险基因型(如基因型16和18)是大多数宫颈癌的罪魁祸首和肛门癌,并在HNSCC的肿瘤发生发挥作用,比例变化,根据区域流行病学数据2。
有几种工具可用于检测 HPV 感染。由于高危HPV感染导致病毒致癌蛋白E6和E73的表达,E6和E7转录本的检测被广泛认为是主动HPV感染鉴定4的黄金标准。HPV RNA CISH 可在 FFPE 样本上执行,这些样本很容易从患有各种 HPV 相关疾病的患者身上获得。其性能在子宫颈、肛门和阴道的鳞状宫内肿瘤中,以及宫颈、肛门和上呼吸道的侵入性鳞状细胞癌中进行了评估:其灵敏度达到98%以上在HPVDNA聚合酶链反应(PCR)阳性病例中。这略好于p16免疫染色(93%)和HPVDNA原位杂交(DNA ISH:97%),这是更常用的。在另一组57例鳞状细胞癌(SCC)中,与HPV DNA ISH相比,HPV RNA CISH具有更好的敏感性(100%对88%)。和特异性(87%对74%)6 。
P16免疫染色是一种间接标记,反映细胞周期中断,这可能是由HPV感染4,7引起(但不完全) 引起的。这种具有成本效益的测试具有良好的灵敏度和负预测值,被美国病理学家学院(CAP)和国际联盟推荐为奥述林克斯癌症(OPC)高危HPV感染的代用品标志物。癌症控制(UICC)8.
虽然本文只关注HNSCC中HPV的检测,但HPV RNA CISH在涉及HPV感染的各种其他条件下与临床相关。例如,这种技术可以提高诊断的低级鳞状颅内病变的宫颈(LSIL,以前称为宫颈内皮瘤,1级[CIN1])的形态不明确的病例9的诊断的准确性。关于口管SCC,HPV RNA CISH允许识别HPV相关SCC,在最近第八版《头颈癌TNM分类》(国际癌症联盟)中,该SCC与HPV无关的口管SCC有别于HPV。控制 [UICC])10.由于HPV相关SCC表现出更好的预后,更长的生存期和增强的放疗和化疗敏感性比HPV无关SCC11,12,13,HPV感染的检测可能会影响病人管理14,15.此外,HPV RNA CISH可用于诊断HPV相关多表皮癌,其信号高于HPVDNACISH16。几个多变量分析表明,E6 和 E7 转录的检测与奥法林格 SCC 总体7、15、17、18 和p16阳性性食管SCC19、20的子组。
在这里,我们介绍在FFPE幻灯片上执行的手动HPV RNA CISH的协议,使用从制造商获得的工具包。

<table>
	<tbody>
		<tr>
			<td>Hematoxylin solution, Gill No. 1</td>
			<td>Merck</td>
			<td>GHS132</td>
			<td></td>
		</tr>
		<tr>
			<td>HybEZ Oven (110v)</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>321710</td>
			<td></td>
		</tr>
		<tr>
			<td>HybEZ slide rack</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>300104</td>
			<td></td>
		</tr>
		<tr>
			<td>ImmEdge Hydrophobic Barrier Pen</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>310018</td>
			<td></td>
		</tr>
		<tr>
			<td>RNAscope 2.5 HD Detection Reagents-BROWN</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>322310</td>
			<td>This kit includes amplification reagents AMP1, AMP2, AMP3, AMP4, AMP5 and AMP6, and detection reagents DAB-A and DAB-B</td>
		</tr>
		<tr>
			<td>RNAscope 3-Plex Negative Control Probe</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>320871</td>
			<td>DAPB</td>
		</tr>
		<tr>
			<td>RNAscope 3-Plex Positive Control Probe</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>320861</td>
			<td>PPIB</td>
		</tr>
		<tr>
			<td>RNAscope H202 &#38; Protease Plus Reagent</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>322330</td>
			<td>Hydrogen Peroxyde x2 and Protease Plus x 1</td>
		</tr>
		<tr>
			<td>RNAscope Probe- HPV16/18</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>311121</td>
			<td></td>
		</tr>
		<tr>
			<td>RNAscope Target Retrieval Reagents</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>322000</td>
			<td></td>
		</tr>
		<tr>
			<td>RNAscope Wash Buffer Reagents</td>
			<td>Advanced Cell Diagnostics Inc.</td>
			<td>310091</td>
			<td>Wash Buffer 50X x4</td>
		</tr>
	</tbody>
</table>

chromogenic in situ hybridization as a tool for hpv-related head and neck cancer diagnosis

人皮球瘤病毒 (HPV) 感染是食管鳞状细胞癌 (OPSCC) 亚型的主要危险因素,该病毒往往与与酒精和烟草相关的 OPSCC 更好的结果相关。HPV病毒RNA的原位杂交(CISH)可实现对致癌蛋白E6和E7病毒转录本的半定量评价,并具有良好的空间分辨率的原位可视化。此技术允许通过肿瘤HPV感染细胞中的HPV转录可视化来诊断活动感染。这种技术的一个优点是避免来自肿瘤附近的非肿瘤HPV感染细胞的污染。总体而言,其良好的诊断性能被认为是主动HPV感染鉴定的黄金标准。由于E6和E7病毒蛋白与细胞蛋白pRb和p3的相互作用是强制性的细胞转化,HPV RNA CISH在功能上相关,并敏锐地反映活性致源性HPV感染。这种技术在临床上也相关,因为"低"或"高"HPV转录水平有助于识别HPV相关p16阳性头颈部癌症患者中的两个预后组。在这里,我们介绍在正式固定石蜡嵌入 (FFPE) 幻灯片上执行的手动 HPV RNA CISH 的协议,该幻灯片使用从制造商处获得的工具包。RNA原位杂交也可以用荧光导光(RNA FISH)进行,而不是染色体性揭示。它也可以与传统的免疫染色相结合。

如此所述,在头颈部鳞状细胞癌中,在细胞质或肿瘤细胞核中存在棕色双核状染色的情况下,可被视为阳性病例。在大多数研究中,信号被认为是"正"或"未检测到"14。信号半量化的方法已经报告,但团队之间缺乏标准化。例如,在一些研究中,信号被评分为 1+,每个肿瘤细胞 1+3 个点,每个肿瘤细胞 4+9 个点 2+,或者每个肿瘤细胞610 个或更多点 3+;在事后分析中,只考虑了2+和3+?...

Watch this Scientific Journal Video about Chromogenic In Situ Hybridization as a Tool for HPV-Related Head and Neck Cancer Diagnosis at JoVE.com

Chromogenic In Situ Hybridization as a Tool for HPV-Related Head and Neck Cancer Diagnosis

人皮疹病毒(HPV)RNA原位染色体杂交被认为是肿瘤内活性人皮疹病毒感染检测的黄金标准之一。它允许通过定位和半定量评估其信号来可视化HPV E6-E7 mRNA表达。

原位杂交的色原性杂交是HPV相关头颈部癌症诊断的工具

Human papillomavirus (HPV) infection is a major risk factor for a subtype of oropharyngeal squamous cell carcinoma (OPSCC), which tends to be associated ...

HPV RNA CISH 允许检测活跃的人类教皇瘤病毒感染。由于原位杂交目标HPV RNA，这种技术允许病毒的活性转录的可视化。具有精确的空间分辨率和良好的诊断性能。HPV 的检测可能支持诊断几种与 HPV 相关的病变，如口咽鳞状细胞癌或宫颈肿瘤。它也可能影响预后。制备缓冲液和反染色试剂后，在大型烧杯中制备一个 x 靶量检索试剂，将 70 毫升 10 倍靶量检索试剂添加到 630 毫升蒸馏水中。将烧杯放在带磁力搅拌器的加热板上，用铝箔盖住。在100摄氏度下煮沸其内容物10至15分钟，确保煮沸时间不超过30分钟。要阻止幻灯片上的过氧化物酶活性，将以前去甲酸并放置在幻灯片架中的组织部分，将四到六滴（刚好够覆盖样品）的过氧化物氧化氢添加到每个幻灯片中，并在室温下孵育 10 分钟。在室温下用蒸馏水清洗两次滑梯两分钟。要打破RNA组织部分的RNA组织边界，首先用爪从沸腾的铝箔中去除铝箔xTRR，然后停止搅拌。缓慢而非常小心地浸入滑架。再次用铝箔盖住烧杯，孵育15分钟。使用爪立即将热滑架转移到蒸馏水浴中，然后洗涤两分钟。然后用新鲜的100%乙醇清洗幻灯片两分钟，让它们在室温下干燥两分钟。接下来，使用疏水屏障笔在每个样品周围绘制一个屏障，并让它干燥至少五分钟。对于蛋白酶消化，将幻灯片放在湿度控制托盘中，并添加每个样品约四滴蛋白酶 Plus。用盖子盖住托盘，在 40 摄氏度下将其插入杂交烤箱 30 分钟。从烤箱中取出托盘，然后取下滑架。一次工作一个幻灯片，快速清除多余的液体，将幻灯片放在浸在装满蒸馏水的染色盘中的幻灯片架中。在室温下用蒸馏水清洗两次，持续搅拌。要开始混合，请轻点幻灯片以去除多余的液体，并将其放回幻灯片架中。从烤箱中取出预热的HPV探针，并添加大约四滴，以完全覆盖每个部分。用盖子盖住托盘，在 40 摄氏度下将其插入烤箱两小时。完成孵育后，从烤箱中取出托盘并拆下滑架。一次一张幻灯片，快速去除多余的液体，将幻灯片放在浸入一个填充一个 x 洗涤缓冲液的染色盘中的幻灯片中，在室温下用不断的搅拌方式清洗幻灯片两分钟，然后用新鲜的一个 x 洗涤缓冲液重复。轻点并轻拂以清除幻灯片中多余的液体，然后将它们再次放在幻灯片架中。每节添加大约四滴室温 AMP1，以完全覆盖每个部分。用盖子盖住托盘，在 40 摄氏度下将其插入烤箱 30 分钟。从烤箱中取出托盘后，拆下滑架。一次工作一个幻灯片，快速去除多余的液体，将幻灯片放在浸有一个 x 洗液缓冲液的染色盘中，在室温下用不断搅拌洗涤两分钟，然后用新鲜的一个 x 洗涤缓冲液重复。在适当尺寸的管中分配相同数量的 DAB-A 和 DAB-B 溶液滴，使每个部分约 120 微升 DAB 基板和涡流。从幻灯片架上取下每张幻灯片，然后轻点以去除多余的液体，然后放回幻灯片架中。将 DAB 混合物的大约 120 微升移液到每个组织部分，确保覆盖这些部分。在室温下孵育10分钟，然后进行反污。将每杯板条放置一滴安装介质，然后将板条放在幻灯片上，让其风干。几个小时后，使用手稿中描述的光学显微镜进行评估。如这里描述的头部和颈部鳞状细胞癌，在细胞质或肿瘤细胞核中出现棕色双核染色，定义的阳性情况。结果分为两个分数，RNA CISH高和低分，观察20倍目标。RNA CISH评分染色被认为是低，当观察到低于50%的肿瘤细胞，并覆盖不到80%的细胞表面。另一方面，RNA CISH得分高，当观察到超过50%的染色癌细胞，或染色表面超过80%以上的肿瘤细胞。人们不应该让幻灯片干涸杂交。别忘了预热探头。对于每个样品，执行阳性和阴性控制，以评估RNA的质量。

Research

JoVE Journal

Genetics

Detection of Copy Number Alterations Using Single Cell Sequencing

单细胞测序拷贝数改变的检测

Detection of genomic changes at single cell resolution is important for characterizing genetic heterogeneity and evolution in normal tissues, cancers, and ...

科研

遗传学

Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice

Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization mFISH and Spectral Karyotyping SKY in Irradiated Mice

通过多色荧光跨染色体畸变稳定的检测<em&gt;原位</em&gt;杂交（渔业部）和光谱核型分析（SKY）的照射小鼠

Ionizing radiation (IR) induces numerous stable and unstable chromosomal aberrations. Unstable aberrations, where chromosome morphology is substantially ...

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

CRISPR/Cas9 和 SsODN 在人细胞中催化点突变修复功能的标准方法研究

Combinatorial gene editing using CRISPR/Cas9 and single-stranded oligonucleotides is an effective strategy for the correction of single-base point ...

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification

高分辨率荧光原位杂交在果蝇胚胎和组织中使用 Tyramide 信号放大

In our efforts to determine the patterns of expression and subcellular localization of Drosophila RNAs on a genome-wide basis, and in a variety of ...

Profiling DNA Replication Timing Using Zebrafish as an In Vivo Model System

Profiling DNA Replication Timing Using Zebrafish as an In Vivo Model System

使用斑马鱼作为体内模型系统分析 DNA 复制定时

DNA replication timing is an important cellular characteristic, exhibiting significant relationships with chromatin structure, transcription, and DNA ...

Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice

Canalostomy 为成人和新生小鼠内耳局部药物传递的外科治疗方法

Local delivery of therapeutic drugs into the inner ear is a promising therapy for inner ear diseases. Injection through semicircular canals (canalostomy) ...

Visualization of Microbiota in Tick Guts by Whole-mount In Situ Hybridization

Visualization of Microbiota in Tick Guts by Whole-mount In Situ Hybridization

全挂载原位杂交法可视化蜱内脏微生物群

Infectious diseases transmitted by arthropod vectors continue to pose a significant threat to human health worldwide. The pathogens causing these ...

Single Molecule Fluorescence In Situ Hybridization (smFISH) Analysis in Budding Yeast Vegetative Growth and Meiosis

Single Molecule Fluorescence In Situ Hybridization smFISH Analysis in Budding Yeast Vegetative Growth and Meiosis

单分子荧光原位杂交 (smFISH) 在发芽酵母营养生长和减数分裂中的分析

Single molecule fluorescence in situ hybridization (smFISH) is a powerful technique to study gene expression in single cells due to its ability to detect ...

Quantitative Analysis of Alternative Pre-mRNA Splicing in Mouse Brain Sections Using RNA In Situ Hybridization Assay

Quantitative Analysis of Alternative Pre-mRNA Splicing in Mouse Brain Sections Using RNA In Situ Hybridization Assay

RNA原位杂交法定量分析小鼠脑切片中的替代前 mRNA 拼接方法

Alternative splicing (AS) occurs in more than 90% of human genes. The expression pattern of an alternatively spliced exon is often regulated in a cell ...

Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization

Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization

原位杂交法在小鼠心脏切片组织特异 miRNA 表达谱分析中的应用

micro-RNAs (miRNAs) are single-stranded RNA transcripts that bind to messenger RNAs (mRNAs) and inhibit their translation or promote their degradation. To ...