Name: single-cell multiplexed fluorescence imaging to visualize viral nucleic acids and proteins and monitor hiv, htlv, hbv, hcv, zika virus, and influenza infection
Uploaded: 2020-10-29T14:00:00
Description: Watch this Scientific Journal Video about Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infectio

这项工作全部或部分得到了美国国立卫生研究院的支持（R01 AI121315，R01 AI146017，R01 AI148382，R01 AI120860，R37 AI076119和U54 AI150472）。我们感谢Raymond F. Schinazi博士和Sadie Amichai提供感染甲型流感病毒的细胞。

1.盖玻片或腔室载玻片上的种子细胞（悬浮液 与 贴壁细胞）（提供的方案使用盖玻片）。
<ol><li>悬浮细胞的接种<ol><li>通过首先将盖玻片在乙醇（EtOH）中孵育5分钟（灭菌盖玻片并去除任何残留物）来制备聚-D-赖氨酸（PDL）包被的盖玻片（促进悬浮细胞粘附到盖玻片上）。然后在磷酸盐缓冲盐水（PBS）中洗涤2x，然后在室温（RT）下将盖玻片在PDL（20μg/ mL）中孵育30分钟（分钟）。</li>
		<li>...

<ol>
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P., Moffitt, J. R., Zhuang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplexed+detection+of+RNA+using+MERFISH+and+branched+DNA+amplification.">Multiplexed detection of RNA using MERFISH and branched DNA amplification.</a> Scientific Reports. 9 (1), 7721 (2019).</li><li>Player, A. N., Shen, L. P., Kenny, D., Antao, V. P., Kolberg, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-copy+gene+detection+using+branched+DNA+(bDNA)+in+situ+hybridization.">Single-copy gene detection using branched DNA (bDNA) in situ hybridization.</a> Journal of Histochemistry and Cytochemistry. 49 (5), 603-612 (2001).</li><li>Battich, N., Stoeger, T., Pelkmans, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image-based+transcriptomics+in+thousands+of+single+human+cells+at+single-molecule+resolution.">Image-based transcriptomics in thousands of single human cells at single-molecule resolution.</a> Nature Methods. 10 (11), 1127-1133 (2013).</li><li>Bushnell, 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=ProbeDesigner:+for+the+design+of+probesets+for+branched+DNA+(bDNA)+signal+amplification+assays.">ProbeDesigner: for the design of probesets for branched DNA (bDNA) signal amplification assays.</a> Bioinformatics. 15 (5), 348-355 (1999).</li><li>Stultz, R. D., Cenker, J. J., McDonald, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+HIV-1+Genomic+DNA+from+Entry+through+Productive+Infection.">Imaging HIV-1 Genomic DNA from Entry through Productive Infection.</a> Journal of Virology. 91 (9), (2017).</li><li>Brown, 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+nuclear+proteins+together+with+transcribed+and+inactive+genes+in+structurally+preserved+cells.">Visualizing nuclear proteins together with transcribed and inactive genes in structurally preserved cells.</a> Methods. 26 (1), 10-18 (2002).</li><li>Francis, 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=HIV-1+replication+complexes+accumulate+in+nuclear+speckles+and+integrate+into+speckle-associated+genomic+domains.">HIV-1 replication complexes accumulate in nuclear speckles and integrate into speckle-associated genomic domains.</a> Nature Communications. 11 (1), 3505 (2020).</li><li>Dharan, A., Bachmann, N., Talley, S., Zwikelmaier, V., Campbell, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+pore+blockade+reveals+that+HIV-1+completes+reverse+transcription+and+uncoating+in+the+nucleus.">Nuclear pore blockade reveals that HIV-1 completes reverse transcription and uncoating in the nucleus.</a> Nature Microbiology. 5 (9), 1088-1095 (2020).</li></ol>

同时可视化 RNA、DNA 和蛋白质通常需要广泛的优化。两种常用的方法是5-乙炔基-2-脱氧尿苷（EdU）标记和DNA FISH。EdU标记已被应用于同时可视化病毒DNA和蛋白质，因为EdU被掺入新生DNA中，随后通过点击化学 用 含叠氮化物的荧光染料标记。因此，EdU标记可用于监测DNA病毒的天然病毒复制动力学或具有DNA模板的病毒进行复制12。然而，EdU标记的一个缺点是，在分裂细胞中，复制...

虽然数以千计的商业抗体可用于通过常规免疫染色方法特异性标记蛋白质，并且虽然融合蛋白可以使用光优化荧光标签进行工程设计以跟踪样品中的多种蛋白质1，但蛋白质的显微可视化通常与传统的DNA荧光原位杂交（FISH）不兼容2.使用基于荧光的方法同时可视化DNA、RNA和蛋白质的技术限制阻碍了对病毒复制的深入了解。在感染过程中跟踪病毒核酸和蛋白质使病毒学家能够可视化病毒复制和组装的基本过程3，4，5，6。
我们开发了一种成像方法，即基于d na，rNA和Protein的成像方法m ultiplex immunofluorescent cell（MICDDRP）3，该方法利用支链DNA（bDNA）原位技术来提高核酸检测的灵敏度7，8，9.此外，该方法利用成对探针来增强特异性。bDNA序列特异性探针使用分支前置放大器和放大器DNA产生强烈的局部信号，改进了以前的杂交方法，该方法依赖于靶向DNA9中的重复区域。临床环境中的受感染细胞通常不含丰富的病毒遗传物质，为诊断环境中荧光核酸检测的灵敏方法提供了商品。通过RNAscope7和ViewRNA10等方法实现bDNA技术的商业化填补了这一空白。bDNA荧光成像的灵敏度在细胞生物学中也具有重要的用途，可以检测细胞培养模型中稀缺的核酸种类。灵敏度的巨大提高使基于bDNA的成像方法适用于研究病毒。然而，一个潜在的缺点是这些方法专注于可视化RNA或RNA和蛋白质。所有复制细胞和许多病毒在其复制周期中都具有DNA基因组或形成DNA，因此非常需要能够对RNA和DNA以及蛋白质进行成像的方法。
在MICDDRP协议中，我们使用RNAscope方法进行bDNA FISH以检测病毒核酸，并修饰7。该方案的主要修改之一是化学固定后蛋白酶处理的优化。蛋白酶处理有助于去除与核酸结合的蛋白质，从而提高探针杂交效率。蛋白酶处理后用支链寡核苷酸探针孵育。应用bDNA探针后，洗涤样品，随后与信号前置放大器和放大器DNA一起孵育。多重原位杂交（ISH），标记多个基因靶标，需要具有不同颜色通道的靶标探针进行光谱分化7。用DNA扩音器孵育后进行免疫荧光（IF）。
bDNA ISH通过放大靶标特异性信号来改善信噪比，并减少非特异性杂交事件的背景噪声7，11。靶探针是使用公开可用的软件程序设计的，这些软件程序预测非特异性杂交事件的概率，并计算探针-目标杂交7，11的熔解温度（Tm）。靶探针包含一个与靶DNA/RNA序列互补的18至25碱基区域、一个间隔序列和一个14个碱基的尾部序列。一对靶探针，每个探针具有不同的尾序列，杂交到目标区域（跨越~50个碱基）。两个尾序列形成前置放大器探针的杂交位点，其中包含20个放大器探针的结合位点，此外还包含20个标记探针的结合位点。例如，核酸分子上的一千碱基（kb）区域被20个探针对靶向，从而创建一个分子支架，用于与前置放大器、放大器和标记探针进行顺序杂交。因此，这可以使每个核酸分子的理论产量达到8000个荧光标记，从而能够检测单个分子，并且比传统的FISH方法7有了巨大的改进（bDNA信号扩增示意图见图1A）。要为多路复用 ISH 设置探针，每个目标探针必须位于不同的颜色通道（C1、C2 或 C3）中。这些具有不同颜色通道的目标探针具有不同的14碱基尾序列。这些尾部序列会将不同的信号放大器与不同的荧光探针结合，从而实现跨多个目标的光谱分化。在所提出的方案中，步骤9中的表4提供了有关荧光标记靶探针的更多信息。此外，图2和图3提供了我们如何选择合适的放大器4-FL（A，B或C）（荧光探针和最终杂交步骤）的示例，以实现HIV-1和HTLV-1感染后多个病毒核酸靶标的特异性荧光标记。
我们已经展示了同时对RNA，DNA和蛋白质进行荧光可视化的几种应用，以高时空分辨率3，4，5观察病毒复制的关键阶段。例如，病毒RNA，细胞质和核DNA以及蛋白质的同时单细胞可视化使我们能够可视化HIV-1感染期间的关键事件，包括在核进入和前病毒DNA3整合之前跟踪细胞质中含有核心的RNA。此外，我们还应用MICDDRP来表征宿主因子和药物治疗对病毒感染和复制的影响4，5。在Ukah等人中，我们跟踪了用不同潜伏逆转剂处理的潜伏期细胞模型中HIV-1转录的再激活，以可视化HIV转录和潜伏期逆转4。此外，MICDDRP可以让我们可视化与小分子治疗或宿主因子限制引起的抗病毒抑制相关的表型变化。作为我们方法稳健性和广泛适用性的概念证明，我们已经证明，我们可以使用我们的方案的修改来有效地标记病毒核酸，以跟踪感染，不仅在人类免疫缺陷病毒（HIV）-1，而且在人类嗜T淋巴细胞病毒（HTLV）-1，乙型肝炎病毒（HBV），丙型肝炎病毒（HCV）， 寨卡病毒（ZIKV）和甲型流感病毒（IAV）。由于HIV-1生命周期由病毒DNA和RNA物种组成，因此我们已经根据HIV-1复制动力学对MICDDRP进行了大部分优化。然而，此外，我们已经证明，我们可以跟踪ZIKV，IAV，HBV和HCV等病毒中或两种感觉（+）和反义（-）链的不同病毒RNA转录本的合成，以监测病毒转录和复制3，4，5，6。这些研究旨在提高对几种病毒过程的机制理解，并作为将这种荧光成像技术应用于各种细胞模型的指南。

<table>
	<tbody>
		<tr>
			<td>4% PFA</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>50% dextran sulfate</td>
			<td>Amreso</td>
			<td>198</td>
			<td>For DNA hybridization buffer</td>
		</tr>
		<tr>
			<td>50X wash buffer</td>
			<td>ACD Bio</td>
			<td>320058</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well plates</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier 1-FL</td>
			<td>ACD Bio</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier 2-FL</td>
			<td>ACD Bio</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier 3-FL</td>
			<td>ACD Bio</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier 4-FL</td>
			<td>ACD Bio</td>
			<td></td>
			<td>Consult Amp-4 table in protocol</td>
		</tr>
		<tr>
			<td>Anti-HCV NS5a antibody</td>
			<td>Abcam</td>
			<td>ab13833</td>
			<td>Mouse monoclonal; works with HCV genotypes 1a, 1b, 3, and 4</td>
		</tr>
		<tr>
			<td>Anti-HIV-1 p24 monoclonal antibody</td>
			<td>NIH AIDS Reagent Program</td>
			<td>3537</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Mov10 antibody</td>
			<td>Abcam</td>
			<td>ab80613</td>
			<td>Rabbit polyclonal</td>
		</tr>
		<tr>
			<td>Anti-PB1 antibody</td>
			<td>GeneTex</td>
			<td>GTX125923</td>
			<td>Antibody against flu protein</td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td></td>
			<td></td>
			<td>Blocking reagent for immunostaining</td>
		</tr>
		<tr>
			<td>Cell media with supplements</td>
			<td></td>
			<td></td>
			<td>Media appropriate for cell model</td>
		</tr>
		<tr>
			<td>Coverslips</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>ACD Bio</td>
			<td></td>
			<td>Nuclear stain (RNAscope kit from ACD Bio)</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s phosphate buffered saline (1X PBS)</td>
			<td>Gibco</td>
			<td>14190250</td>
			<td>No calcium and magnesium</td>
		</tr>
		<tr>
			<td>Ethylene carbonate</td>
			<td>Sigma</td>
			<td>E26258</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum (FBS)</td>
			<td></td>
			<td></td>
			<td>Use specific FBS based on what serum secondary antibody was raised in (e.g goat FBS)</td>
		</tr>
		<tr>
			<td>Fisherbrand colorfrost plus microscope slides</td>
			<td>Fisher Scientific</td>
			<td>12-550-17/18/19</td>
			<td>Precleaned</td>
		</tr>
		<tr>
			<td>HCV-GT2a-sense-C2 probe</td>
			<td>ACD Bio</td>
			<td>441371</td>
			<td>HCV(+) sense RNA probe</td>
		</tr>
		<tr>
			<td>HIV-gagpol-C1</td>
			<td>ACD Bio</td>
			<td>317701</td>
			<td>HIV-1 cDNA probe</td>
		</tr>
		<tr>
			<td>HIV-nongagpol-C3</td>
			<td>ACD Bio</td>
			<td>317711-C</td>
			<td>HIV-1 RNA probe</td>
		</tr>
		<tr>
			<td>HybEZ hybridization oven</td>
			<td>ACD Bio</td>
			<td>321710/321720</td>
			<td></td>
		</tr>
		<tr>
			<td>ImmEdge hydrophobic barrier pen</td>
			<td>Vector Laboratories</td>
			<td>H-4000</td>
			<td></td>
		</tr>
		<tr>
			<td>Nail polish</td>
			<td></td>
			<td></td>
			<td>For immobolizing coverslip to slide prior to protease treatment</td>
		</tr>
		<tr>
			<td>Nuclease free water</td>
			<td>Ambion</td>
			<td>AM9937</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-d-lysine (PDL)</td>
			<td></td>
			<td></td>
			<td>Coat coverslips in 20 &#181;g/mL of PDL for 30 minutes</td>
		</tr>
		<tr>
			<td>Probe diluent</td>
			<td>ACD Bio</td>
			<td>300041</td>
			<td>For diluting RNA C2 or C3 probes</td>
		</tr>
		<tr>
			<td>Prolong gold antifade</td>
			<td>Invitrogen</td>
			<td>P36930</td>
			<td></td>
		</tr>
		<tr>
			<td>Protease III</td>
			<td>ACD Bio</td>
			<td>322337</td>
			<td></td>
		</tr>
		<tr>
			<td>RNAscope&#174;&#160;Probe- V-Influenza-H1N1-H5N1-NP</td>
			<td>ACD Bio</td>
			<td>436221</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase A</td>
			<td>Qiagen</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Secondary antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Slides</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td></td>
			<td></td>
			<td>For DNA hybridization buffer</td>
		</tr>
		<tr>
			<td>Sodium citrate, pH 6.2</td>
			<td></td>
			<td></td>
			<td>For DNA hybridization buffer</td>
		</tr>
		<tr>
			<td>Tween-20</td>
			<td></td>
			<td></td>
			<td>For DNA hybridization buffer and PBS-T</td>
		</tr>
		<tr>
			<td>V-HBV-GTD</td>
			<td>ACD Bio</td>
			<td>441351</td>
			<td>Total HBV RNA</td>
		</tr>
		<tr>
			<td>V-HBV-GTD-01-C2</td>
			<td>ACD Bio</td>
			<td>465531-C2</td>
			<td>HBV pgRNA probe</td>
		</tr>
		<tr>
			<td>V-HCV-GT2a probe</td>
			<td>ACD Bio</td>
			<td>441361</td>
			<td>HCV(-) sense RNA probe</td>
		</tr>
		<tr>
			<td>V-HTLV-HBZ-sense-C3</td>
			<td>ACD Bio</td>
			<td>495071-C3</td>
			<td>HTLV-1 (-) sense RNA probe targetting HBZ</td>
		</tr>
		<tr>
			<td>V-HTLV1-GAG-C2</td>
			<td>ACD Bio</td>
			<td>495051-C2</td>
			<td>HTLV-1 DNA probe</td>
		</tr>
		<tr>
			<td>V-HTLV1-GAG-POL-sense</td>
			<td>ACD Bio</td>
			<td>495061</td>
			<td>HTLV-1 (+) sense RNA probe</td>
		</tr>
		<tr>
			<td>V-Influenza-H1N1-H5N1-NP</td>
			<td>ACD Bio</td>
			<td>436221</td>
			<td>IAV RNA probe</td>
		</tr>
		<tr>
			<td>V-ZIKA-pp-O2</td>
			<td>ACD Bio</td>
			<td>464531</td>
			<td>Zika(+) sense RNA probe</td>
		</tr>
		<tr>
			<td>V-ZIKA-pp-O2-sense-C2</td>
			<td>ACD Bio</td>
			<td>478731-C2</td>
			<td>Zika(-) sense RNA probe</td>
		</tr>
	</tbody>
</table>

single-cell multiplexed fluorescence imaging to visualize viral nucleic acids and proteins and monitor hiv, htlv, hbv, hcv, zika virus, and influenza infection

由于缺乏能够灵敏和同时标记病毒核酸和蛋白质的强大原位杂交（ISH）技术，捕获病毒的动态复制和组装过程受到了阻碍。传统的DNA荧光原位杂交（FISH）方法通常与免疫染色不兼容。因此，我们开发了一种成像方法 MICDDRP （multiplex immunofluorescent c ell-basedd etection of D NA， RNA和 Protein），该方法能够同时进行DNA，RNA和蛋白质的单细胞可视化。与传统的DNA鱼相比，MIDDRP采用支链DNA（bDNA）ISH技术，可显著提高寡核苷酸探针的灵敏度和检测。MICDDRP的微小修饰能够同时对病毒蛋白进行成像，同时具有不同链的核酸（RNA或DNA）。我们已经应用这些协议来研究多种病毒病原体的生命周期，包括人类免疫缺陷病毒（HIV）-1，人类嗜T淋巴细胞病毒（HTLV）-1，乙型肝炎病毒（HBV），丙型肝炎病毒（HCV），寨卡病毒（ZKV）和甲型流感病毒（IAV）。我们证明了我们可以有效地标记各种病毒中的病毒核酸和蛋白质。这些研究可以为我们提供对多种病毒系统的机理理解，此外，还可以作为在广泛的细胞系统中应用DNA，RNA和蛋白质的多重荧光成像的模板。

MICDDRP的示意图如图 1所示。标记DNA和RNA之后进行免疫染色。支化放大器的使用增加了信号，允许检测单个核酸分子。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61843/61843fig1v5.jpg"> 图 1：MICDDRP 和分步工作流程示意图。 （A）bDNA信号通过分支前置放大器和放大器DNA放大，以增强对病毒DNA（1）和RNA物种?...

Watch this Scientific Journal Video about Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infectio

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

这里介绍的是荧光成像方法的 协议，multiplex immunofluorescent cELL 基于d etection of DNA， RNA和 Protein（MICDDRP），该方法能够同时对不同类型和链的病毒蛋白和核酸进行荧光单细胞可视化。这种方法可以应用于各种系统。

单细胞多重荧光成像，可可视化病毒核酸和蛋白质，并监测 HIV、HTLV、HBV、HCV、寨卡病毒和流感感染

Capturing the dynamic replication and assembly processes of viruses has been hindered by the lack of robust in situ hybridization (ISH) technologies that ...

Research

JoVE Journal

Biology

Purification and Visualization of Influenza A Viral Ribonucleoprotein Complexes

A型流感病毒的核蛋白复合物的分离纯化及可视化

The influenza A viral genome consists of eight negative-sense, single stranded RNA molecules, individually packed with multiple copies of the influenza A ...

科研

生物学

A Technique to Simultaneously Visualize Virus-Specific CD8+ T Cells and Virus-Infected Cells In situ

A Technique to Simultaneously Visualize Virus-Specific CD8+ T Cells and Virus-Infected Cells In situ

一种技术，同时可视化病毒特异性CD8 + T细胞和病毒感染的细胞原位</em

The numbers and locations of virus-specific CD8+ T cells relative to the numbers and locations of their infected cell targets is thought to be critical in ...

NanoDrop Microvolume Quantitation of Nucleic Acids

NanoDrop微量核酸的定量

Biomolecular assays are continually being developed that use progressively smaller amounts of material, often precluding the use of conventional ...

Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer

使用微量的生物规格的纳米分光光度计核酸和蛋白的浓度测定

Nucleic Acid quantitation procedures have advanced significantly in the last three decades.  More and more, molecular biologists require consistent ...

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

化学阻断抗体芯片复用高通量的复杂样品中特定蛋白糖基化的剖析

 In this study, we describe an effective protocol for use in a multiplexed high-throughput antibody microarray with glycan binding protein detection that ...

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

核酸，蛋白质与甲基转移和辅酶类似物序列特异性标签

S-Adenosyl-l-methionine (AdoMet or SAM)-dependent methyltransferases (MTase) catalyze the transfer of the activated methyl group from AdoMet to specific ...

Filtration Isolation of Nucleic Acids:  A Simple and Rapid DNA Extraction Method

核酸的分离过滤：一种简单快速的DNA提取方法

FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad ...

Functional Imaging of Viral Transcription Factories Using 3D Fluorescence Microscopy

基于3D 荧光显微镜的病毒转录工厂功能成像

It is well known that spatial and temporal regulation of genes is an integral part of governing proper gene expression. Consequently, it is invaluable to ...

Single Cell Micro-aspiration as an Alternative Strategy to Fluorescence-activated Cell Sorting for Giant Virus Mixture Separation

单细胞微吸气作为荧光激活细胞分类的替代策略，用于巨型病毒混合物分离

During the amoeba co-culture process, more than one virus may be isolated in a single well. We previously solved this issue by end point dilution and/or ...

Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

探索病毒感染时的m6A和m5C表继转录组：HIV示例

The role of RNA modifications in biological processes has been the focus of an increasing number of studies in the last few years and is known nowadays as ...