Name: visualization of sars-cov-2 using immuno rna-fluorescence in situ hybridization
Uploaded: 2020-12-23T14:00:00
Description: Watch this Scientific Journal Video about Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization at JoVE.com

这项工作得到了科学和高等教育部对SARS-CoV-2研究的支持，并得到了国家科学中心的赠款（向K.P.和UMO-2018/30/E/NZ1/00874赠款给A.K.-P.）。

1. 缓冲区准备
<ol><li>对于 500 mL 的 2 倍 PHEM 缓冲器，将 18.14 克管道拉辛 -N，N+-二（2-乙烷硫酸）（PIPES），6.5 克 4-（2-羟基苯甲酸） 1-管道丙烷硫酸（HEPES）、3.8克乙二醇四乙酸（EGTA）和0.99克硫酸镁（MgSO4）。用蒸馏水（dH2O）使体积达到~400毫升，搅拌，然后使用10M氢氧化钾（KOH）或氢氧化钠（NAOH）将pH调整到7.0。使最终音量达到 500 mL，然后拆分为 50 mL 的别名。储存在-20°...

<ol>
	<li>Choi, H. M. 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=Third-generation+in+situ+hybridization+chain+reaction:+multiplexed,+quantitative,+sensitive,+versatile,+robust.">Third-generation in situ hybridization chain reaction: multiplexed, quantitative, sensitive, versatile, robust.</a> Development. 145, (2018).</li><li>Milewska, 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=Replication+of+severe+acute+respiratory+syndrome+coronavirus+2+in+human+respiratory+epithelium.">Replication of severe acute respiratory syndrome coronavirus 2 in human respiratory epithelium.</a> Journal of Virology. 94, (2020).</li><li>Banach, B. 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G. 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=ACE2+expression+is+increased+in+the+lungs+of+patients+with+comorbidities+associated+with+severe+COVID-19.">ACE2 expression is increased in the lungs of patients with comorbidities associated with severe COVID-19.</a> Journal of Infectious Diseases. 222 (4), 556-563 (2020).</li><li>Verdecchia, P., Cavallini, C., Spanevello, A., Angeli, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pivotal+link+between+ACE2+deficiency+and+SARS-CoV-2+infection.">The pivotal link between ACE2 deficiency and SARS-CoV-2 infection.</a> European Journal of Internal Medicine. 76, 14-20 (2020).</li><li>Gordon, D. E., 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+SARS-CoV-2+protein+interaction+map+reveals+targets+for+drug+repurposing.">A SARS-CoV-2 protein interaction map reveals targets for drug repurposing.</a> Nature. 583, 459-468 (2020).</li><li>Ferron, F., Decroly, E., Selisko, B., Canard, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+viral+RNA+capping+machinery+as+a+target+for+antiviral+drugs.">The viral RNA capping machinery as a target for antiviral drugs.</a> Antiviral Research. 96 (1), 21-31 (2012).</li><li>Mehta, S., Jeffrey, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+receptors+and+signaling:+epigenetic+factors+in+the+regulation+of+innate+immunity.">Beyond receptors and signaling: epigenetic factors in the regulation of innate immunity.</a> Immunology and Cell Biology. 93 (3), 233-244 (2015).</li><li>Poppe, M., 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+NF-kappaB-dependent+and+-independent+transcriptome+and+chromatin+landscapes+of+human+coronavirus+229E-infected+cells.">The NF-kappaB-dependent and -independent transcriptome and chromatin landscapes of human coronavirus 229E-infected cells.</a> PLoS Pathogens. 13, 1006286 (2017).</li><li>Schafer, A., Baric, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+landscape+during+coronavirus+infection.">Epigenetic landscape during coronavirus infection.</a> Pathogens. 6 (1), 8 (2017).</li><li>Carletti, 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=Viral+priming+of+cell+intrinsic+innate+antiviral+signaling+by+the+unfolded+protein+response.">Viral priming of cell intrinsic innate antiviral signaling by the unfolded protein response.</a> Nature Communications. 10, 3889 (2019).</li><li>Heil, 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=Species-specific+recognition+of+single-stranded+RNA+via+toll-like+receptor+7+and+8.">Species-specific recognition of single-stranded RNA via toll-like receptor 7 and 8.</a> Science. 303 (5663), 1526-1529 (2004).</li><li>Sze, 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=Host+restriction+factor+SAMHD1+limits+human+T+cell+leukemia+virus+type+1+infection+of+monocytes+via+STING-mediated+apoptosis.">Host restriction factor SAMHD1 limits human T cell leukemia virus type 1 infection of monocytes via STING-mediated apoptosis.</a> Cell Host and Microbe. 14 (4), 422-434 (2013).</li><li>Kikkert, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innate+immune+evasion+by+human+respiratory+RNA+viruses.">Innate immune evasion by human respiratory RNA viruses.</a> Journal of Innate Immunity. 12, 4-20 (2020).</li><li>Kennedy, E. M., 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=Posttranscriptional+m(6)A+editing+of+HIV-1+mRNAs+enhances+viral+gene+expression.">Posttranscriptional m(6)A editing of HIV-1 mRNAs enhances viral gene expression.</a> Cell Host and Microbe. 22 (6), 830 (2017).</li><li>Fulcher, M. L., Gabriel, S., Burns, K. A., Yankaskas, J. R., Randell, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Well-differentiated+human+airway+epithelial+cell+cultures.">Well-differentiated human airway epithelial cell cultures.</a> Human Cell Culture Protocols in Methods in Molecular Medicine. 107, 183-206 (2005).</li><li>Milewska, A., Owczarek, K., Szczepanski, A., Pyrc, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+coronavirus+entry+into+cells.">Visualizing coronavirus entry into cells.</a> Methods in Molecular Biology. 2203, 241-261 (2020).</li></ol>

免疫-RNA-FISH是RNA和细胞蛋白双重染色的可靠方法。在这里，描述了一个经过修改的免疫-RNA-FISH协议，允许检测SARS-CoV-2RNA和细胞蛋白的细胞系和HAE培养物。此协议可适用于不同的细胞模型，包括细胞单层或特定组织。该方法依赖于由适当的探头本地化启动的HCR的概念。接下来，使用分发启动器探头开始放大荧光标记放大器的信号，当使用共聚焦显微镜观察时，可产生极小到无背景荧光。放大器可以...

SARS-CoV-2是一种新型的人类β可罗纳病毒，于2019年底出现，几个月后引发了前所未有的大流行。由于该病毒是科学的新生病毒，因此其大部分生物学及其对宿主细胞的影响仍不得而知。因此，如果要了解病毒细胞和组织在感染过程中的三胞头，其基本的生物学特征及其对宿主的影响就很重要。使用多种技术来检查病毒宿主之间的相互作用，包括生化、生物和物理检测。原位杂交是一种常用方法，它采用标记的互补DNA、RNA或改性核酸探针，该探针定位到细胞或组织中的特定DNA或RNA序列。
一种新的RNA荧光原位杂交（RNA-FISH）方法已经开发出来，它结合了修改，通过HCR1放大信号与噪声比来增加灵敏度。HCR 允许在单细胞级别上研究 RNA 本地化。由于其特异性、灵敏度和分辨率高，这种方法不仅对基础科学研究有用，而且对应用项目（如诊断）也很有用。最近，该方法的可行性被证明用于检测SARS-CoV-2RNA本地化为完全分化的3D人类气道上皮（HAE）培养物2中的附属细胞。HAE文化是用于研究"自然感染"微环境3、4背景下病毒感染的最先进工具之一。
包括SARS-CoV-2在内的几份关于人类冠状病毒（HCoV）的报告强调了表观遗传学对HCoV感染和病理生理学（在5中回顾）的重要性，例如编码血管紧张素转换酶2（ACE-2）受体6、7的基因的甲基化模式。有趣的是，质谱筛查发现了几个与SARS-CoV-2蛋白体8相互作用的表观遗传因素。更具体地说，非结构蛋白 5 （NSP5） 与表观遗传调节器、高石脱乙酰酶 2 结合，催化不活跃 NSP5 （C145A） 与 tRNA 甲基转移酶 1 （24） 相互作用。此外，NSP16甲基转移酶活性被甲基转移酶抑制剂西内芬金9阻断。然而，这些表观遗传因素在COVID-19中的确切作用尚不清楚。HCoV的复制发生在受感染细胞的细胞质中，并触发由表观遗传修饰调节的炎症反应10。
例如，HCoV-229E微调核因子-卡帕B信号，并通过在某些区域增加H3K36和H4K5的乙酰化，对宿主细胞色度图进行深刻重新编程。中东呼吸系统综合症相关冠状病毒感染增加了H3K27me3的水平，并耗尽了H3K4me3在特定干扰素敏感基因12子集的促进区域。此外，病毒RNA触发细胞免疫反应，如病毒13，逆转录病毒14，15和冠状病毒16证明。病毒RNA上的表观遗传标记可能起到细胞传感器识别的作用，如人类免疫缺陷病毒-1 RNA17的m7A甲基化所示。然而，问题依然存在：SARS-CoV-2RNA对免疫反应有何影响，是否涉及表观遗传标记？
在这里，介绍了一种优化的RNA-FISH方法，以及细胞系和3D组织（完全分化HAE）的免疫荧光分析。虽然细胞学方法，如FISH和免疫荧光，被广泛使用，这种基于HCR的新一代原位杂交方法从未用于病毒检测（除了在最近的出版物）2。一般来说，免疫染色和鱼氧化物需要以下步骤：渗透，使探针或抗体穿透：固定其中细胞材料固定和保存;检测其中应用抗体或核酸探针;最后，安装样品进行可视化。
虽然现有协议具有这些通用功能，但它们在所涉及的参数方面存在显著差异。在这里，一个优化的，简单的，免疫-RNA-FISH协议已经描述检测SARS-CoV-2RNA在HAE培养和维罗细胞。该技术包括以下步骤：（1） 用甲醛固定细胞：（2） 用洗涤剂或甲醇（MeOH） 渗透：（3） 通过一系列分级的 MeOH 解决方案（仅限 HAE 文化）进行补液：（4） 检测：（五）利用HCR技术检测SARS-CoV-2RNA的放大：（六）免疫学：和（7）在共焦显微镜下成像。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td >Equipment</td>
			<td ></td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >Confocal Microscope LSM 880</td>
			<td >ZEISS </td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Grant Bio, Mini Rocker- Shaker</td>
			<td >Fisher Scientific</td>
			<td>12965501</td>
			<td></td>
		</tr>
		<tr >
			<td >Incubator Galaxy170R</td>
			<td >New Brunswick</td>
			<td>CO170R-230-1000</td>
			<td></td>
		</tr>
		<tr >
			<td >Thermomixer Comfort</td>
			<td >Eppendorf</td>
			<td>5355 000 011</td>
			<td></td>
		</tr>
		<tr >
			<td>Materials</td>
			<td></td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >15 mm x 15 mm NO. 1 coverslips</td>
			<td >LabSolute</td>
			<td >7695022</td>
			<td ></td>
		</tr>
		<tr >
			<td >1.5 mL tubes</td>
			<td >FL-MEDICAL</td>
			<td >5.350.023.053</td>
			<td ></td>
		</tr>
		<tr >
			<td >12-well plate</td>
			<td >TTP</td>
			<td >92412</td>
			<td ></td>
		</tr>
		<tr >
			<td >Conical centrifuge tube</td>
			<td >Sarstedt</td>
			<td >5.332.547.254</td>
			<td ></td>
		</tr>
		<tr >
			<td >parafilm</td>
			<td >Sigma</td>
			<td >P7793-1EA</td>
			<td ></td>
		</tr>
		<tr >
			<td >serological pipets</td>
			<td >VWR Collection</td>
			<td >612-5523P, 612-5827P</td>
			<td ></td>
		</tr>
		<tr >
			<td >slide glass</td>
			<td >PTH CHEMLAND</td>
			<td >04-296.202.03</td>
			<td ></td>
		</tr>
		<tr >
			<td >Transwell ThinCerts</td>
			<td >Grainer bio-one</td>
			<td >665641</td>
			<td ></td>
		</tr>
		<tr >
			<td >Reagents</td>
			<td ></td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >Alexa fluorophore 488-conjugated secondary antibodies </td>
			<td>Invitrogen</td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >&#946;5-tubulin </td>
			<td>Santa Cruz Biotechnology</td>
			<td > sc-134234</td>
			<td ></td>
		</tr>
		<tr >
			<td >DAPI</td>
			<td >Thermo Scientific</td>
			<td >D1306</td>
			<td ></td>
		</tr>
		<tr >
			<td >Disodium phosphate</td>
			<td >Sigma</td>
			<td >S51136-500G</td>
			<td ></td>
		</tr>
		<tr >
			<td >EGTA</td>
			<td >BioShop</td>
			<td >EGT101.25</td>
			<td ></td>
		</tr>
		<tr >
			<td >HCR Amplification Buffer</td>
			<td >Molecular Instruments, Inc.</td>
			<td >BAM01522</td>
			<td >Buffer can be also prepared doi:10.1242/dev.165753: Supplementary information</td>
		</tr>
		<tr >
			<td >HCR amplifier B1-h1 Alexa Fluor 647</td>
			<td >Molecular Instruments, Inc.</td>
			<td >S013922</td>
			<td ></td>
		</tr>
		<tr >
			<td >HCR amplifier B1-h2 Alexa Fluor 647</td>
			<td >Molecular Instruments, Inc.</td>
			<td >S012522</td>
			<td ></td>
		</tr>
		<tr >
			<td >HCR Probe Hybridization Buffer</td>
			<td >Molecular Instruments, Inc.</td>
			<td >BPH03821</td>
			<td >Buffer can be also prepared doi:10.1242/dev.165753: Supplementary information</td>
		</tr>
		<tr >
			<td >HCR probe set for SARS-CoV-2 Ncapsid</td>
			<td >Molecular Instruments, Inc.</td>
			<td >PRE134</td>
			<td></td>
		</tr>
		<tr >
			<td >HCR Probe Wash Buffer</td>
			<td >Molecular Instruments, Inc.</td>
			<td >BPW01522</td>
			<td >Buffer can be also prepared doi:10.1242/dev.165753: Supplementary information</td>
		</tr>
		<tr >
			<td >HEPES</td>
			<td >BioShop</td>
			<td >HEP001.100</td>
			<td ></td>
		</tr>
		<tr >
			<td >Magnesium sulfate heptahydrate</td>
			<td >Sigma</td>
			<td >63138-250G</td>
			<td ></td>
		</tr>
		<tr >
			<td >Methanol</td>
			<td >Sigma</td>
			<td >32213-1L-M</td>
			<td ></td>
		</tr>
		<tr >
			<td >Monopotassium phosphate</td>
			<td >Sigma</td>
			<td >P5655-100G</td>
			<td ></td>
		</tr>
		<tr >
			<td >Paraformaldehyde</td>
			<td >Sigma</td>
			<td >P6148-1KG</td>
			<td ></td>
		</tr>
		<tr >
			<td >PIPES</td>
			<td >BioShop</td>
			<td >PIP666.100</td>
			<td ></td>
		</tr>
		<tr >
			<td >Potassium Chloride</td>
			<td >Sigma</td>
			<td >P5405-250G</td>
			<td ></td>
		</tr>
		<tr >
			<td >Prolong Diamond Antifade Mounting Medium</td>
			<td >Invitrogen</td>
			<td >P36970</td>
			<td ></td>
		</tr>
		<tr >
			<td >Sodium Chloride</td>
			<td >BioShop</td>
			<td >SOD001.5</td>
			<td ></td>
		</tr>
		<tr >
			<td >Trisodium Citrate 2-hydrate</td>
			<td >POCH</td>
			<td > 6132-04-3</td>
			<td ></td>
		</tr>
		<tr >
			<td >Tween-20</td>
			<td >BioShop</td>
			<td >TWN580.500</td>
			<td ></td>
		</tr>
		<tr >
			<td ></td>
			<td >Software</td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Fluorescence Spectraviewer</td>
			<td ></td>
			<td></td>
			<td>Modeling spectral parameters</td>
		</tr>
		<tr >
			<td >ImageJ Fiji</td>
			<td ></td>
			<td ></td>
			<td >Acquiring and processing z-stack images</td>
		</tr>
	</tbody>
</table>

visualization of sars-cov-2 using immuno rna-fluorescence in situ hybridization

本手稿提供了原位杂交链反应 （HCR） 与免疫荧光的协议，可直观地将严重急性呼吸系统综合征冠状病毒 2 （SARS-CoV-2） RNA 在细胞系中和人类气道上皮的三维 （3D） 培养物中。该方法允许高度具体和敏感的病毒RNA可视化，依靠HCR启动的探针本地化。拆分启动器探头通过荧光标记的放大器帮助放大信号，导致共焦显微镜中可忽略不计的背景荧光。用不同的荧光染料标记放大器有助于同时识别各种目标。这反过来又使组织中的感染图谱能够更好地了解单细胞水平的病毒发病机能和复制。将这种方法与免疫荧光相结合，可以促进更好地了解宿主-病毒相互作用，包括宿主表观基因组和免疫反应通路的交替。由于敏感和特定的HCR技术，此协议也可以用作诊断工具。同样重要的是要记住，该技术可能很容易修改，以便能够检测任何RNA，包括将来可能出现的非编码RNA和RNA病毒。

本手稿中描述的免疫-RNA-FISH协议使用两个蜂窝系统执行：维罗细胞系和3D HAE培养物。两个蜂窝模型的主要步骤都显示在 表 2 中。RNA-FISH 在 HAE 培养中可视化 SARS-CoV-2 的协议包括组织样本的典型步骤，即通过一系列 MeOH-PBS 和 0.1% Tween 解决方案，以 100% MeOH 渗透和补液。免疫荧光是在RNA-FISH完成之后进行的。获取和处理了 Zstack 图像。
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Watch this Scientific Journal Video about Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization at JoVE.com

Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization

在这里，我们描述了一个简单的方法，结合RNA荧光原位杂交（RNA-FISH）与免疫荧光可视化严重急性呼吸系统综合症冠状病毒2（SARS-CoV-2）RNA。该协议可以增加对SARS-CoV-2 RNA-主机在单细胞水平上的分子特征的理解。

使用免疫RNA-荧光原位杂交对SARS-CoV-2进行可视化

This manuscript provides a protocol for in situ hybridization chain reaction (HCR) coupled with immunofluorescence to visualize severe acute respiratory ...

Research

JoVE Journal

Immunology and Infection

Fluorescent in situ Hybridization on Mitotic Chromosomes of Mosquitoes

Fluorescent in situ Hybridization on Mitotic Chromosomes of Mosquitoes

荧<em在原位</em&gt;蚊虫的有丝分裂染色体上的杂交

Fluorescent in situ hybridization (FISH) is a technique routinely used by many laboratories to determine the chromosomal position of DNA and RNA probes. ...

科研

免疫与感染

Visualization of Candida albicans in the Murine Gastrointestinal Tract Using Fluorescent In Situ Hybridization

Visualization of Candida albicans in the Murine Gastrointestinal Tract Using Fluorescent In Situ Hybridization

使用荧光原位杂交对毛尿胃肠道中的念珠菌的可视化

Candida albicans is a fungal component of the gut microbiota in humans and many other mammals. Although C. albicans does not cause symptoms in most ...

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection

使用假病毒感染的高通量荧光成像检测SARS-CoV-2中和抗体

As the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to evolve, it has become evident that the ...

A Rapid, Multiplex Dual Reporter IgG and IgM SARS-CoV-2 Neutralization Assay for a Multiplexed Bead-Based Flow Analysis System

基于多路复用珠子的流量分析系统的快速、多路双记者 IgG 和 Igm SARS-CoV-2 中和检测

The COVID-19 pandemic has underscored the need for rapid high-throughput methods for sensitive and specific serological detection of infection with novel ...

Detection of SARS-CoV-2 Receptor-Binding Domain Antibody using a HiBiT-Based Bioreporter

使用基于HiBiT的生物报告器检测SARS-CoV-2受体结合域抗体

The emergence of the COVID-19 pandemic has increased the need for better serological detection methods to determine the epidemiologic impact of severe ...

Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging

Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging

通过荧光 原位 杂交、凝集素染色和成像可视化肠道微生物群与宿主的相互作用

Measuring the localization of microbes within their in vivo context is an essential step in revealing the functional relationships between the microbiota ...

Live Imaging and Quantification of Viral Infection in K18 hACE2 Transgenic Mice Using Reporter-Expressing Recombinant SARS-CoV-2

使用报告基因表达重组SARS-CoV-2对K18 hACE2转基因小鼠的病毒感染进行实时成像和定量

The coronavirus disease 2019 (COVID-19) pandemic has been caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To date, SARS-CoV-2 has ...

Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source Pipetting Robots

利用开源移液机器人的高效SARS-CoV-2定量逆转录酶PCR唾液诊断策略

The emergence of the recent SARS-CoV-2 global health crisis introduced key challenges for epidemiological research and clinical testing. Characterized by ...

RNA Fluorescence in situ Hybridization (FISH) to Visualize Microbial Colonization and Infection in Caenorhabditis elegans Intestines

RNA Fluorescence in situ Hybridization FISH to Visualize Microbial Colonization and Infection in Caenorhabditis elegans Intestines

RNA荧光 原位 杂交（FISH）可视化 秀丽隐杆线虫 肠道中的微生物定植和感染

The intestines of wild Caenorhabditis nematodes are inhabited by a variety of microorganisms, including gut microbiome bacteria and pathogens, such as ...

Quantifying Vaccine-Induced Neutralizing Antibodies in SARS-CoV-2 Variants

Application of Ha-CoV-2 Pseudovirus for Rapid Quantification of SARS-CoV-2 Variants and Neutralizing Antibodies

Author Spotlight: A Pseudotype Virus System for Assessing Omicron Subvariants and Neutralizing Antibodies in SARS-CoV-2 Research

The coronavirus disease 2019 pandemic (COVID-19) has highlighted the need for rapid assays to accurately measure the infectivity of emerging SARS-CoV-2 ...