Name: quantification of antibody-dependent enhancement of the zika virus in primary human cells
Uploaded: 2019-01-18T13:30:00
Description: Watch this Scientific Journal Video about Quantification of Antibody-dependent Enhancement of the Zika Virus in Primary Human Cells at JoVE.com

这项工作得到了 1R21AI129881-01 (对 t. m. c.)、国家新兴传染病实验室和波士顿大学医学院启动资金的慷慨支持。

本研究中使用的血清样本来自哥伦比亚一个群体的人类参与者。项目收集工作得到了潘普洛纳大学 (南美洲哥伦比亚) 和洛斯波蒂奥斯医院8号内部审查委员会的批准。这些样本是匿名提供的, 调查人员无法获得病人信息。对血清样本进行了 denv 血清型检测。这些样本进一步证实在体外中和中和 denv 感染。为了进行控制, 使用了来自美国健康人 (hc) 的血清样本。
<p class="jove...

<ol>
	<li>Hayes, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zika+virus+outside+Africa.">Zika virus outside Africa.</a> Emerging Infectious Diseases. 15, 1347-1350 (2009).</li><li>Grard, G., 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=Zika+virus+in+Gabon+(Central+Africa)+-+2007:+a+new+threat+from+Aedes+albopictus.">Zika virus in Gabon (Central Africa) - 2007: a new threat from Aedes albopictus.</a> PLoS Neglected Tropical Diseases. 8 (2), 2681 (2014).</li><li>Fauci, A. S., Morens, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zika+Virus+in+the+Americas--Yet+Another+Arbovirus+Threat.">Zika Virus in the Americas--Yet Another Arbovirus Threat.</a> New England Journal of Medicine. 374 (7), 601-604 (2016).</li><li>Hawkes, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+the+Infectivity+of+Arboviruses+by+Specific+Antisera+Produced+in+Domestic+Fowls.">Enhancement of the Infectivity of Arboviruses by Specific Antisera Produced in Domestic Fowls.</a> Australian Journal of Experimental Biology and Medical Science. 42, 465-482 (1964).</li><li>Musso, D., Gubler, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zika+Virus.">Zika Virus.</a> Clinical Microbiology Reviews. 29 (3), 487-524 (2016).</li><li>Vaughn, D. 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=Dengue+viremia+titer,+antibody+response+pattern,+and+virus+serotype+correlate+with+disease+severity.">Dengue viremia titer, antibody response pattern, and virus serotype correlate with disease severity.</a> The Journal of Infectious Diseases. 181 (1), 2-9 (2000).</li><li>Khandia, R., 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=Modulation+of+Dengue/Zika+Virus+Pathogenicity+by+Antibody-Dependent+Enhancement+and+Strategies+to+Protect+Against+Enhancement+in+Zika+Virus+Infection.">Modulation of Dengue/Zika Virus Pathogenicity by Antibody-Dependent Enhancement and Strategies to Protect Against Enhancement in Zika Virus Infection.</a> Frontiers of Immunology. 9, 597 (2018).</li><li>Londono-Renteria, B., 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=A+relevant+in+vitro+human+model+for+the+study+of+Zika+virus+antibody-dependent+enhancement.">A relevant in vitro human model for the study of Zika virus antibody-dependent enhancement.</a> Journal of General Virology. 98 (7), 1702-1712 (2017).</li><li>Ganger, M. T., Dietz, G. D., Ewing, S. 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+common+base+method+for+analysis+of+qPCR+data+and+the+application+of+simple+blocking+in+qPCR+experiments.">A common base method for analysis of qPCR data and the application of simple blocking in qPCR experiments.</a> BMC Bioinformatics. 18 (1), 534 (2017).</li><li>Renn, L. 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=High-throughput+quantitative+real-time+RT-PCR+assay+for+determining+expression+profiles+of+types+I+and+III+interferon+subtypes.">High-throughput quantitative real-time RT-PCR assay for determining expression profiles of types I and III interferon subtypes.</a> Journal of Visualized Experiments. (97), e52650 (2015).</li><li>McArthur, M. 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=Dengue+vaccines:+recent+developments,+ongoing+challenges+and+current+candidates.">Dengue vaccines: recent developments, ongoing challenges and current candidates.</a> Expert Review of Vaccines. 12 (8), 933-953 (2013).</li><li>Priyamvada, L., 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=Humoral+cross-reactivity+between+Zika+and+dengue+viruses:+implications+for+protection+and+pathology.">Humoral cross-reactivity between Zika and dengue viruses: implications for protection and pathology.</a> Emerging Microbes and Infections. 6 (5), 33 (2017).</li><li>Dai, L., 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=Molecular+basis+of+antibody-mediated+neutralization+and+protection+against+flavivirus.">Molecular basis of antibody-mediated neutralization and protection against flavivirus.</a> IUBMB Life. 68 (10), 783-791 (2016).</li><li>Dai, L., 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=Structures+of+the+Zika+Virus+Envelope+Protein+and+Its+Complex+with+a+Flavivirus+Broadly+Protective+Antibody.">Structures of the Zika Virus Envelope Protein and Its Complex with a Flavivirus Broadly Protective Antibody.</a> Cell Host &amp; Microbe. 19 (5), 696-704 (2016).</li><li>Sirohi, D., 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+3.8+A+resolution+cryo-EM+structure+of+Zika+virus.">The 3.8 A resolution cryo-EM structure of Zika virus.</a> Science. 352 (6284), 467-470 (2016).</li><li>George, 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=Prior+Exposure+to+Zika+Virus+Significantly+Enhances+Peak+Dengue-2+Viremia+in+Rhesus+Macaques.">Prior Exposure to Zika Virus Significantly Enhances Peak Dengue-2 Viremia in Rhesus Macaques.</a> Scientific Reports. 7 (1), 10498 (2017).</li><li>Morens, D. M., Halstead, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+antibody-dependent+infection+enhancement+of+four+dengue+virus+serotypes+by+monoclonal+and+polyclonal+antibodies.">Measurement of antibody-dependent infection enhancement of four dengue virus serotypes by monoclonal and polyclonal antibodies.</a> Journal of General Virology. 71, 2909-2914 (1990).</li><li>Dejnirattisai, 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=Cross-reacting+antibodies+enhance+dengue+virus+infection+in+humans.">Cross-reacting antibodies enhance dengue virus infection in humans.</a> Science. 328 (5979), 745-748 (2010).</li><li>Priyamvada, L., 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+antibody+responses+after+dengue+virus+infection+are+highly+cross-reactive+to+Zika+virus.">Human antibody responses after dengue virus infection are highly cross-reactive to Zika virus.</a> Proceedings of the National Academy of Sciences of the United States of America. 113 (28), 7852-7857 (2016).</li><li>Charles, A. S., Christofferson, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utility+of+a+Dengue-Derived+Monoclonal+Antibody+to+Enhance+Zika+Infection+In+Vitro.">Utility of a Dengue-Derived Monoclonal Antibody to Enhance Zika Infection In Vitro.</a> PLoS Currents. 8, (2016).</li><li>Swanstrom, 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=Dengue+Virus+Envelope+Dimer+Epitope+Monoclonal+Antibodies+Isolated+from+Dengue+Patients+Are+Protective+against+Zika+Virus.">Dengue Virus Envelope Dimer Epitope Monoclonal Antibodies Isolated from Dengue Patients Are Protective against Zika Virus.</a> MBio. 7 (4), (2016).</li></ol>

denv 抗体的交叉反应导致其他 denv 血清型的 ade 阻碍了有效疫苗的开发11。zikv 属于同一家族, flaviviridae, 与其他黄病毒, 特别是 denv12有相当多的同源性。zikv 和 denv 中和抗体的主要目标是包络蛋白, 它在这两种病毒13、14、15之间共享一个非常高的结构和四元序列同源。已经证明, 前免疫要么 denv ...

在蚊子传播的病毒性疾病中, 寨卡感染是临床上最重要的疾病之一。感染是由黄病毒 zikv 引起的, 在大多数情况下, 黄病毒使用埃及伊蚊作为其主要载体1,2。然而, 有研究报告白纹伊蚊是一些 zikv疫情3的主要媒介。虽然感染在许多情况下是无症状的, 但最常见的症状是发烧、头痛和肌肉疼痛2。没有治疗或疫苗可用于 zikv 感染, 现有的治疗方法大多是支持性的。最近在南美洲爆发的 zikv 导致了严重的病例, 并使被称为 "小脑2" 的胎儿的神经发育障碍增加了大约20倍。由于南美洲是 denv 和西尼罗河病毒等几种弓形虫病毒的特有地区, 因此调查先前对其他黄病毒的免疫力是否在 zikv 感染和疾病的严重程度中起着重要作用至关重要。
随着年龄的增长, 病毒已经进化出不同的策略, 以增加其感染的机会, 以接管宿主细胞机械, 并抑制抗病毒反应。其中最引人入胜的是利用宿主免疫前抗体的病毒, 以提高其复制与现象 ade 4.在 denv 的所有四个血清型中, ade 都得到了很好的研究和证明, 以增加病毒滴度和疾病结果5, 6,7.在先前的体外研究中, 我们已经证明了 zikv 复制的显著增强, 由于已有的 denv 免疫在原代人类免疫细胞8。我们还演示了一种相关的体外方法来量化 denv 先前存在的抗体增强初级细胞 zikv 复制的能力。
我们开发的协议使用在 tcid-50 或斑块还原中和试验 (prnt) 检测中和测试的人类血清样本, 以及在生物相关的细胞或从 zikv 可以感染的组织中提取的细胞中使用 zikv。

<table border="0" cellpadding="0" cellspacing="0" style="width:773px;" width="773">
	<colgroup>
		<col />
		<col span="2" />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">Fetal Bovine Serum&#160;</td>
			<td style="width:119px;">GEMINI</td>
			<td style="width:119px;">100-106</td>
			<td style="width:277px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">iCycler&#160;</td>
			<td style="width:119px;">BioRad</td>
			<td style="width:119px;">785BR02188</td>
			<td>Model No. CFX96 Optics Module</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">Microfuge 18 Centrifuge</td>
			<td style="width:119px;">Beckman Coulter&#160;</td>
			<td style="width:119px;">367160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">Nanodrop-1000</td>
			<td style="width:119px;">Thermoscientific&#160;</td>
			<td style="width:119px;">1072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">Quantifast SYBR-One step RT-PCR kit&#160;</td>
			<td style="width:119px;">Qiagen&#160;</td>
			<td style="width:119px;">204154</td>
			<td>Used for 1 step RT-qPCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">RNeasy RNA Isolation Kit&#160;</td>
			<td style="width:119px;">Qiagen&#160;</td>
			<td style="width:119px;">74106</td>
			<td>Used for RNA extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:259px;">RPMI-medium&#160;</td>
			<td style="width:119px;">Gibco</td>
			<td style="width:119px;">11875093</td>
			<td></td>
		</tr>
	</tbody>
</table>

quantification of antibody-dependent enhancement of the zika virus in primary human cells

最近出现的黄病毒寨卡和神经并发症, 如桂林-巴雷综合征和婴儿小脑, 带来了严重的公共安全问题。在危险因素中, 抗体依赖性增强 (ade) 构成了最严重的威胁, 因为最近寨卡病毒 (zikv) 的重新出现主要发生在人群接触过的地区, 处于对其他密切相关的人群的免疫前状态病毒, 尤其是登革热病毒 (denv)。在这里, 我们描述了一个程序, 量化人类血清抗体对初级人类细胞或细胞系的 zikv 感染的影响。

在图 1中, 有一个逐步的图表, 说明了执行 ade 协议所涉及的所有步骤。这是一个示意图, 显示了整个过程的 zikv 由于预先存在的对 denv 的免疫力。图 2显示了人类血清样本如何分为三组: denv 感染确认样本被称为 denv 感染组, denv 抗体确认样本被称为 denv 暴露组, 健康没有 denv 中和抗体或 rna 的个体血清称为健康控制 (hc) 组。(<strong c...

Watch this Scientific Journal Video about Quantification of Antibody-dependent Enhancement of the Zika Virus in Primary Human Cells at JoVE.com

Quantification of Antibody-dependent Enhancement of the Zika Virus in Primary Human Cells

我们描述了一种方法来评估现有的免疫对登革热病毒感染的影响, 使用人血清, 原代人类细胞, 和感染定量定量定量定量定量定量定量定量定量定量定量定量定量的定量。

原代人细胞中寨卡病毒抗体依赖性增强的定量研究

The recent emergence of the flavivirus Zika and neurological complications, such as Guillain-Barr&#233; syndrome and microcephaly in infants, has brought ...

该方法可用于评估登革热病毒的原发免疫对寨卡病毒感染的影响，使用实际的人类患者血清和原发性人体免疫细胞。使用实际的人类患者血清样本和人原细胞，通过免疫前血清和抗体，更深入地了解寨卡病毒的抗体依赖性增强。这种方法提供了对患者血液中抗体增强其他人类细胞中病毒感染的能力的洞察。重要的是要确保病毒感染水平足够高，足以检测，但足够低，不压倒细胞或干扰数据解释。首先在无菌、平底96井的盘子里，每井100微升细胞培养中播种三倍十至第四细胞。当所有细胞都镀上时，将板放在37摄氏度和5%的二氧化碳培养箱中，并在无血清介质中连续稀释10倍的解冻人体血清样本。在将稀释添加到病毒溶液和细胞之前，请确保所有免疫前血清的血清稀释和稀释因子保持不变。接下来，在37摄氏度的水浴中解冻寨卡病毒株，在一到两分钟内将库存迅速转移到冰中。将寨卡病毒的感染等效量 0.1 倍数添加到血清等分中，并将板放在细胞培养箱中一小时，使登革热病毒抗体与 Zika 病毒形成复合物。在孵化结束时，用每井100微升PBS洗涤细胞，并在每井中加入50微升的免疫复合物。经过两个小时的孵育，取出介质，用每井100微升PBS洗两次，以完全去除免疫复合物和任何未附着的病毒。然后给细胞喂100微升，每井细胞培养，辅以10%的胎儿牛血清。然后将细胞返回到细胞培养箱48小时。感染后两天，用每口100微升无菌PBS洗两次细胞，然后向每口井添加250微升细胞解液缓冲液，每口井中加入10%β汞醇。通过刮擦上下移液至少五次，以加快分解过程，将细胞解液转移到无菌标记的 1.5 毫升管中。在每个管中加入70%乙醇的等量，并移液，上下四到五次。当利沙酸盐悬浮液清晰时，将每个溶液转移到两毫升收集管中标记的二氧化硅柱中进行离心。丢弃流式保持同一收集管中的柱，并将 700 微升的洗涤缓冲液 1 添加到每列进行第二次离心。丢弃流式处理后，使用每次洗涤两次的 500 微升洗涤缓冲液再次冲洗柱子。第二次洗涤后，将柱转移到新的两毫升收集管中，进行另一次离心。将 30 微升 42 摄氏度无 RNase 水添加到每个柱的中心进行离心并回收洗净的 RNA。对于定量实时聚合酶链反应或QRTPCR分析，从10微摩尔库存中加入一个基因特异性前进和反向质剂的微升，以及每反应0.25微升的逆转录酶混合物。请注意，识别寨卡病毒遗传物质的底材将检测病毒基因并允许感染定量。接下来，添加 12.5 微升的 SYBR 绿色混合，然后每次反应中加入 25 微升水，在每个 96 井 QPCR 板的井中加入 15 微升主混合。使用主混合时，将 100 纳米RNA样品添加到混合中，并将主混合移液器放入 QRTPCR 板的单个孔中。请注意，样品应在同一 QRTPCR 板上以三次三次运行。然后根据表中概述的参数在定量 PCR 机器上运行样本。单击系统软件中的熔体曲线选项卡以监视熔体曲线。熔体曲线将在特定基因的所有样本中显示单峰，以确认只有一个安培康的存在。为了获得最佳效果，在熔体曲线协议中，在步骤之间使用 0.5 摄氏度的温度增量和 10 秒的最小保持时间。然后单击量化数据选项卡以获取每个样本的定量周期，然后将数据导出到电子表格中进行进一步的定量分析。人血清样本可分为三组：登革热病毒感染确认样本、登革热病毒抗体确认样本、无登革热病毒中和抗体或RNA的健康血清。经过48小时的感染，QRTPCR分析表明，大多数含有登革热病毒血清型的血清一至四抗体能够增强寨卡病毒在不同水平上的复制。寨卡病毒滴答声增加率最高的是接受含有血清素的登革热病毒血清型2和4抗体治疗的巨噬细胞，而血清型1和3显示寨卡病毒的诱导相对较低。重要的是要保持无菌环境，穿适当的个人防护设备，并始终将解冻的病毒血清样本和QPCR试剂放在冰上。该协议可以很容易地修改，以研究抗体依赖增强的其他病毒，如黄热病病毒，登革热病毒，或西尼罗河病毒使用一组患者血清。这项技术对于在arbo病毒或病毒学领域进行未来的抗体依赖性增强研究将非常有用。应时刻使用适当的生物安全二级个人防护设备，所有病毒废物应在处置前 10%漂白中除污染 30 分钟。

Research

JoVE Journal

Immunology and Infection

piggyBac Transposon System Modification of Primary Human T Cells

piggyBac Transposon System Modification of Primary Human T Cells

piggyBac转座子初级人类T细胞系统改造

The piggyBac transposon system is naturally active, originally derived from the cabbage looper moth1,2. This non-viral system is plasmid based, most ...

科研

免疫与感染

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

中性粒细胞分离和分析，以确定他们的淋巴瘤细胞的敏感性对治疗药物的作用

Neutrophils are the most abundant (40% to 75%) type of white blood cells and among the first inflammatory cells to migrate towards the site of ...

Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons

Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons

兹卡病毒感染细胞培养系统和<em&gt;体外</em&gt;干扰素的预防作用

Zika Virus (ZIKV) is an emerging pathogen that is linked to fetal developmental abnormalities such as microcephaly, eye defects, and impaired growth. ZIKV ...

Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells

葡聚糖增强小鼠和人原发性 NK 细胞的慢转导效率

The efficient transduction of specific genes into natural killer (NK) cells has been a major challenge. Successful transductions are critical to defining ...

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone

来自新世界Zika病毒感染性克隆的重组病毒的救援和表征

Infectious cDNA clones allow for genetic manipulation of a virus, thus facilitating work on vaccines, pathogenesis, replication, transmission and viral ...

Zika Virus Specific Diagnostic Epitope Discovery

Zika 病毒特定诊断表位发现

High-density peptide microarrays allow screening of more than six thousand peptides on a single standard microscopy slide. This method can be applied for ...

A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum

A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum

基于流式细胞仪的 Zika 病毒疗养血清测定登革热病毒的体外抗体依赖性

Antibody dependent enhancement of infection has been shown to play a major role in Dengue viral pathogenesis. Traditional assays that measure the capacity ...

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

感染 Ifnar1 小鼠 Immunoprivileged 器官寨卡病毒特异 T 细胞反应的评价

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and ...

Quantification of Efferocytosis by Single-cell Fluorescence Microscopy

单细胞荧光显微镜定量 Efferocytosis 的研究

Studying the regulation of efferocytosis requires methods that are able to accurately quantify the uptake of apoptotic cells and to probe the signaling ...

Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse

从小鼠多个器官分离和定量寨卡病毒

The methods being presented demonstrate laboratory procedures for the isolation of organs from Zika virus infected animals and the quantification of viral ...