Name: quantitative fluorescence in situ hybridization (fish) and immunofluorescence (if) of specific gene products in kshv-infected cells
Uploaded: 2019-08-27T14:30:00
Description: Watch this Scientific Journal Video about Quantitative Fluorescence In Situ Hybridization FISH and Immunofluorescence IF of Specific Gene Products in KSHV-Infected Cells at JoVE.com

我们感谢乔纳森·罗登费尔斯、卡齐米尔兹·泰科夫斯基和约翰娜·威瑟斯就数据分析提供建议。我们还感谢G.海沃德的抗SSB抗体。这项工作得到了国家卫生研究院（TKV）和NIH赠款（CA16038）（JAS）的T32GM007223和T32AI055403的资助。JAS是霍华德·休斯医学研究所的研究员。图1-3和表1经美国微生物学会许可转载，从以下出版物获得知识共享归因许可：瓦莱里、T.K.、威瑟斯、J.B.、安多、斯蒂茨、J.A.卡波西的《萨科马联系》核病毒中的疱疹病毒mRNA积累受病毒DNA复制和病毒非编码多基因化核RNA的影响。病毒学杂志。92 （13）， doi：10.1128/JVI.00220-18， （2018）。

1. 原位荧光（FISH）抗感寡核苷酸，以检测特定的疱疹病毒记录仪
<ol><li>从感兴趣的RNA序列中选择25到40个nt段，并转换为抗感。成功的FISH策略可能包含多达十种或更多不同的反义寡核苷酸。选择序列时，请考虑以下事项：<ol>
<li>如果感兴趣的RNA包含一个独特的重复区域，则利用此功能并设计一个抗感寡核苷酸以靶向重复序列。 注：Tycowski 和同事5提供了此策略的示例，包括河?...

<ol>
	<li>Amen, M. A., Griffiths, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Packaging+of+Non-Coding+RNAs+into+Herpesvirus+Virions:+Comparisons+to+Coding+RNAs.">Packaging of Non-Coding RNAs into Herpesvirus Virions: Comparisons to Coding RNAs.</a> Frontiers in Genetics. 2, 81 (2011).</li><li>Schmid, M., Speiseder, T., Dobner, T., Gonzalez, 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=DNA+virus+replication+compartments.">DNA virus replication compartments.</a> Journal of Virology. 88 (3), 1404-1420 (2014).</li><li>Pawlicki, J. M., Steitz, 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=Primary+microRNA+transcript+retention+at+sites+of+transcription+leads+to+enhanced+microRNA+production.">Primary microRNA transcript retention at sites of transcription leads to enhanced microRNA production.</a> Journal of Cell Biology. 182 (1), 61-76 (2008).</li><li>Borah, S., Darricarrere, N., Darnell, A., Myoung, J., Steitz, 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=A+viral+nuclear+noncoding+RNA+binds+re-localized+poly(A)+binding+protein+and+is+required+for+late+KSHV+gene+expression.">A viral nuclear noncoding RNA binds re-localized poly(A) binding protein and is required for late KSHV gene expression.</a> Public Library of Science Pathogens. 7 (10), e1002300 (2011).</li><li>Tycowski, K. T., Shu, M. D., Borah, S., Shi, M., Steitz, J. 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A., Penman, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+molecular+weight+monodisperse+nuclear+RNA.">Small molecular weight monodisperse nuclear RNA.</a> Journal of Molecular Biology. 38 (3), 289-304 (1968).</li><li>Myoung, J., Ganem, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+a+doxycycline-inducible+KSHV+producer+cell+line+of+endothelial+origin:+maintenance+of+tight+latency+with+efficient+reactivation+upon+induction.">Generation of a doxycycline-inducible KSHV producer cell line of endothelial origin: maintenance of tight latency with efficient reactivation upon induction.</a> Journal of Virology Methods. 174 (1-2), 12-21 (2011).</li><li>Brulois, K. 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=Construction+and+manipulation+of+a+new+Kaposi's+sarcoma-associated+herpesvirus+bacterial+artificial+chromosome+clone.">Construction and manipulation of a new Kaposi's sarcoma-associated herpesvirus bacterial artificial chromosome clone.</a> Journal of Virology. 86 (18), 9708-9720 (2012).</li><li>Sturzl, M., Gaus, D., Dirks, W. G., Ganem, D., Jochmann, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kaposi's+sarcoma-derived+cell+line+SLK+is+not+of+endothelial+origin,+but+is+a+contaminant+from+a+known+renal+carcinoma+cell+line.">Kaposi's sarcoma-derived cell line SLK is not of endothelial origin, but is a contaminant from a known renal carcinoma cell line.</a> International Journal of Cancer. 132 (8), 1954-1958 (2013).</li><li>Chiou, C. 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=Patterns+of+gene+expression+and+a+transactivation+function+exhibited+by+the+vGCR+(ORF74)+chemokine+receptor+protein+of+Kaposi's+sarcoma-associated+herpesvirus.">Patterns of gene expression and a transactivation function exhibited by the vGCR (ORF74) chemokine receptor protein of Kaposi's sarcoma-associated herpesvirus.</a> Journal of Virology. 76 (7), 3421-3439 (2002).</li><li>Cole, R. W., Jinadasa, T., Brown, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+and+interpreting+point+spread+functions+to+determine+confocal+microscope+resolution+and+ensure+quality+control.">Measuring and interpreting point spread functions to determine confocal microscope resolution and ensure quality control.</a> Nature Protocols. 6 (12), 1929-1941 (2011).</li><li>Nakamura, 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=Global+changes+in+Kaposi's+sarcoma-associated+virus+gene+expression+patterns+following+expression+of+a+tetracycline-inducible+Rta+transactivator.">Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator.</a> Journal of Virology. 77 (7), 4205-4220 (2003).</li><li>Majerciak, V., Yamanegi, K., Zheng, Z. 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本报告中描述的方案可以适应不同的细胞类型，包括使用单克隆和多克隆原抗体的IF双RNA FISH和RNA FISH的步骤。虽然制备的幻灯片通常使用共聚焦显微镜进行成像，但经过抗体浓度增加和不同的安装介质的修改后，可以使用 STED（刺激发射消耗）显微镜进行成像。为了增强对单个细胞的分析，使用该协议制备的样品也可以由细胞分拣器或流动细胞仪进行分类、成像和分析，变化不大，如Borah及其<sup c...

在其溶性（活性）阶段，疱疹病毒劫持宿主细胞，导致细胞形态和生物分子的定位变化，以产生病毒。手术的基础是细胞核，其中双链DNA病毒基因组被复制并包装成一个蛋白质外壳，称为辣椒1。首先，病毒表达自己的蛋白质，劫持宿主机制，防止非必需宿主基因的表达，这个过程称为宿主关闭效应。这种活动的大部分被局部化为特定的4°，6-二酰胺-2-phenyinole（DAPI）无核区域称为病毒复制隔间，由宿主和病毒蛋白、RNA和病毒DNA2组成。细胞经过大修，为复制舱提供空间和资源，从而组装病毒辣椒。一旦辣椒离开细胞核，细胞质中如何包裹辣椒，产生膜结合的病毒颗粒，也称为病毒，目前还不清楚。了解宿主和病毒生物分子在溶质阶段中的定位和空间变化，有助于深入了解复制舱的排列、宿主关闭效应、病毒-出口通路和其他与疱疹病毒感染和复制相关的过程。
目前，检测和研究这些变化的最佳方法是分别对免疫荧光（IF）和荧光原位杂交（FISH）受感染细胞中的蛋白质和RNA进行可视化。使用这些技术的时间过程揭示了生物分子在lytic相的关键点或简单的时空数据的定位。FISH 和 IF 补充了其他生化技术，例如抑制细胞过程（例如抑制病毒DNA复制）、RT-qPCR（实时聚合酶链反应）、RNA 测序、北方印迹、质谱法、西方印迹和分析病毒DNA的产生，这可能提供一个更全球性的细胞活动图景。
我们开发了RNA FISH策略来检查来自特定基因的RNA产物，并进行了定量计算特定基因产物核细胞质比的计算分析。从Steitz和同事3，4的早期出版物中修改的样品制备相对容易，可用于粘附细胞和悬浮细胞。该协议还适用于同时使用多个RNA FISH策略（双RNA FISH）或RNA FISH与IF策略。制定具体的 FISH 战略具有挑战性，但概述了提高成功性的建议。如果使用荧光珠和隔间边界的强标记，并且提供对显微图的更多洞察，从而消除观测偏差，则此处描述的数据分析是定量的。详细方案是专为受卡波西肉瘤相关疱疹病毒（KSHV）感染的潜伏细胞和溶血细胞而设计的，可用于未感染的细胞或其他疱疹病毒感染的细胞5。定量方法适用于大多数细胞中核细胞质转移或亚细胞间重新定位的研究。

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			<td height="21" style="height:21px;width:361px;">AlexaFluor594-5-dUTP</td>
			<td style="width:191px;">Life Technologies</td>
			<td style="width:117px;">C1100</td>
			<td style="width:277px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;">anti-DIG FITC&#160;</td>
			<td>Jackson Lab Immunologicals</td>
			<td>200-092-156</td>
			<td style="width:277px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anti-Rabbit Secondary AlexaFluor594 Monoclonal Antibody</td>
			<td>Invitrogen</td>
			<td>A-11037</td>
			<td>Goat</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anti-SSB Antibody</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Ref. Chiou et al. 2002</td>
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		<tr height="21">
			<td height="21" style="height:21px;">BLASTn</td>
			<td>NIH NCBI</td>
			<td>N/A</td>
			<td style="width:277px;">Free Sequence Alignment Software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dextran Sulfate</td>
			<td>Sigma Aldrich</td>
			<td>D8906</td>
			<td style="width:277px;">Molecular Biology Grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DIG-Oligonucleotide Tailing Kit</td>
			<td>Sigma Roche</td>
			<td>#03353583910</td>
			<td style="width:277px;">2nd Gen</td>
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			<td height="42" style="height:42px;">Eight-Chamber Slides</td>
			<td>Nunc Lab Tek II</td>
			<td>#154453</td>
			<td style="width:277px;">Blue seal promotes surface tension but separation by clear gel is also available.&#160;</td>
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			<td height="21" style="height:21px;">Formamide</td>
			<td>Sigma Aldrich</td>
			<td>F9037</td>
			<td style="width:277px;">Molecular Biology Grade</td>
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		<tr height="21">
			<td height="21" style="height:21px;">GAPDH Probes</td>
			<td>Stellaris</td>
			<td>SMF-2019-1</td>
			<td style="width:277px;">Compatible with protocol, Quasar 670</td>
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			<td height="42" style="height:42px;">ImageJ&#160;</td>
			<td>NIH, Bethesda, MD</td>
			<td>N/A</td>
			<td style="width:277px;">Free Image Analysis Software, [http:rsb.info.nih.gov/ij/]</td>
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		<tr height="21">
			<td height="21" style="height:21px;">OligoAnalyzer</td>
			<td>IDT</td>
			<td>N/A</td>
			<td style="width:277px;">Free Oligonucleotide Analyzer&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pcDNA3</td>
			<td>Invitrogen</td>
			<td>A-150228</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pmaxGFP</td>
			<td>Amaxa</td>
			<td>VDF-1012</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Poly L-Lysine</td>
			<td>Sigma Aldrich</td>
			<td>P8920</td>
			<td style="width:277px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Terminal Transferase</td>
			<td>Sigma Roche</td>
			<td>#003333574001</td>
			<td style="width:277px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vanadyl Ribonucleoside Complexes&#160;</td>
			<td>NEB</td>
			<td>S1402S</td>
			<td style="width:277px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Vectashield</td>
			<td>Vector Laboratories, Inc.&#160;</td>
			<td>H-1000</td>
			<td style="width:277px;">DAPI within the mounting media scatters the light and reduces contrast.&#160;</td>
		</tr>
	</tbody>
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quantitative fluorescence in situ hybridization (fish) and immunofluorescence (if) of specific gene products in kshv-infected cells

机械洞察力来自对特定RNA和蛋白质的仔细研究和定量。这些生物分子在特定时间在整个细胞中的相对位置可以通过荧光原位杂交（FISH）和免疫荧光（IF）捕获。在溶性疱疹病毒感染过程中，病毒劫持宿主细胞以优先表达病毒基因，导致细胞形态和生物分子行为发生变化。Lytic 活动以核工厂为中心，称为病毒复制隔间，仅与 FISH 和 IF 一起可辨别。在这里，我们描述了一个适应RNA FISH和IF技术卡波西的肉瘤相关疱疹病毒（KSHV）感染细胞，无论是粘附的还是悬浮的。该方法包括特定抗感寡核苷酸、双RNA FISH、带IF的RNA FISH和荧光强度定量计算的步骤。该协议已成功应用于多种细胞类型、未感染细胞、潜伏细胞、溶酶细胞、时间过程和用抑制剂处理的细胞，以分析来自人类宿主和KSHV.

本手稿中详述的 FISH 和 IF 方法如图1所示，以及按荧光强度线痕迹对结果进行量化。这里给出的结果是半定量的，提供了对定位的洞察，而不是不同荧光污渍的强度的比较，因为实验在幻灯片制备中不包括荧光珠。图1还显示，细胞质和核区域及其比率对于潜伏和溶性KSHV感染细胞是不同的。因此，面积在图2</strong...

Watch this Scientific Journal Video about Quantitative Fluorescence In Situ Hybridization FISH and Immunofluorescence IF of Specific Gene Products in KSHV-Infected Cells at JoVE.com

Quantitative Fluorescence In Situ Hybridization FISH and Immunofluorescence IF of Specific Gene Products in KSHV-Infected Cells

我们描述了一种利用荧光原位杂交（FISH）来可视化被神经感染的人体细胞中的多种疱疹病毒RNA的协议，无论是悬浮的还是粘附的。该协议包括荧光的定量，产生核细胞质比，并可扩展用于同时可视化具有免疫荧光（IF）的宿主和病毒蛋白。

KSHV感染细胞中特定基因产物的定量原位荧光杂交（FISH）和免疫荧光（IF）

Mechanistic insight arrives from careful study and quantification of specific RNAs and proteins. The relative locations of these biomolecules throughout ...

该协议有助于在宿主细胞的上下文中，特别是与疱疹病毒相关的病毒环境中，获得对病毒基因产品的时间调控的定量洞察。这种技术可以帮助解决与宿主和病毒RNA和蛋白质相关的研究问题。此外，该技术与同步生化测定相容。为了将细胞粘附在8个腔室的幻灯片上，将200个固有细胞悬浮液涂抹到无菌的8室滑轨的每个腔室，并允许种子在37摄氏度和5%的二氧化碳下生长12-24小时。在孵育结束时，吸上经细胞和未连接的细胞，并立即在冰上PBS中用预冷却4%甲醛固定细胞30分钟。在固定结束时，用三、五分钟的洗涤用200微升洗涤细胞，每次洗涤温度为4摄氏度的PBS。上次洗涤后，将固定细胞渗透200微升，每井PBS中预冷却0.5%Triton-X，在冰上10分钟。在渗透结束时，小心地取出腔室，而不会裂开幻灯片，然后用预冷的 PBS 冲洗细胞。在4摄氏度下用PBS中预冷却4%BSA的冲洗细胞，在4摄氏度下用适当的多克隆原抗体在PBS中孵育1小时，在4摄氏度。在孵育结束时，用新鲜的PBS清洗细胞三次，用适当的二级抗体孵育细胞，与与鱼检测抗体相容的荧光在4摄氏度下孵育一小时。在演示的三次 PBS 洗涤后，在 PBS 中再次将样品修复为 4%甲醛 10-15 分钟，然后进行第二次渗透。如所证明的。然后用铝箔盖住幻灯片，以保留荧光信号并防止光漂白。在应用45微升杂交溶液之前，用2x盐水柠檬酸钠清洗细胞。在加湿室37摄氏度下一小时后，将蒸馏水加入感兴趣的反义寡核苷酸，使变性量达到10微升。将寡核苷酸在 95 摄氏度下变性 5 分钟，然后每张幻灯片添加 35 微升新鲜杂交溶液。然后在37摄氏度的加湿室中孵育样品10-24小时，用铝箔保护。第二天，用两个，10分钟洗细胞。2X 盐水柠檬酸钠在室温下清洗，随后在 25 摄氏度下用 1X 盐水柠檬酸钠洗涤 10 分钟。上次洗涤后，在PBS中用预冷藏4%甲醛固定细胞，在冰上10-15分钟，然后用新鲜PBS进行三次清洗。接下来，如所证明的，渗透样品，然后用抗二恶英FTC孵育，在PBS中预冷却0.1%BSA，在4摄氏度下孵育1小时。在孵育结束时，用新鲜 PBX 洗涤幻灯片三次，并在 PBS 中用预冷却的 4% 半甲醛固定样品，在 4 摄氏度下 10-15 分钟。在 PBS 中洗三次后，在 PBS 中用每毫升 DAPI 标记 0.4 微克，并在 PBS 中预冷却 0.5%Triton-X 15 分钟，随后在 PBS 中进行三次最终洗涤。安装带荧光珠的幻灯片，作为实验相关且适当的安装介质。用无菌湿巾去除多余的安装介质后，用多层透明指甲油将一个盖玻片密封到每个幻灯片上，并使用荧光显微镜确认实验成功执行。然后，图像可以拍摄到共合显微镜上，在安装后一小时到一周内采集样品的图像，放大倍数为 630 倍。这些具有代表性的 FISH 和免疫荧光结果具有半量化性，有助于深入了解寡核苷酸的定位，而不是比较不同荧光染色的强度，因为这些实验在幻灯片制备中不包括荧光珠。在这些实验中，细胞质和核区域及其比例对于潜伏和细胞解KSHV感染细胞是不同的。如图所示，面积控制在核质比中。当KSHV DNA复制在裂解阶段受到抑制时，KSHV寡核苷酸转录的早期裂解蛋白转向以细胞质为主的定位。然而，KSHV多面体RNA，尽管抑制了病毒DNA复制，却对特定核位点进行局部化。从幻灯片中去除腔室时，注意工具上粘剂残留物很少，并使用乙醇对细胞进行渗透并削弱粘合剂。生物化学测定，如QPCR，西印，高通量测序，可以同时执行，以增加从显微图收集的定量数据。

Research

JoVE Journal

Immunology and Infection

Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells

Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce

Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce

胶粘剂基于磁带的采样和荧光组合原位杂交快速检测沙门氏菌新鲜农产品

This protocol describes a simple approach for adhesive-tape-based sampling of tomato and other fresh produce surfaces, followed by on-tape fluorescence in ...

科研

免疫与感染

Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant

Multivirus特异性T细胞的生成，预防/治疗异基因造血干细胞移植后的病毒感染

Viral infections cause morbidity and mortality in allogeneic hematopoietic stem cell transplant (HSCT) recipients. We and others have successfully ...

Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection

Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization LNA flow-FISH: a Method for Bacterial Small RNA Detection

锁核酸流式细胞仪，荧光原位杂交（LNA流FISH）：细菌小RNA检测方法

Fluorescence in situ hybridization (FISH) is a powerful technique that is used to detect and localize specific nucleic acid sequences in the cellular ...

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

杂交<em&gt;原位</em&gt;和蚊虫唾液腺，卵巢，胚胎

Mosquitoes are vectors for a diverse set of pathogens including arboviruses, protozoan parasites and nematodes.  Investigation of transcripts and gene ...

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. ...

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 ...

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

量<em在体外</em&gt;激活的主人微血管内皮细胞在静态条件下测量的检测，以中性粒细胞粘附

The vascular  endothelium plays an integral part in the inflammatory response. During the  acute phase of inflammation, endothelial cells (ECs) are ...

Fluorescence in situ Hybridizations (FISH) for the Localization of Viruses and Endosymbiotic Bacteria in Plant and Insect Tissues

Fluorescence in situ Hybridizations FISH for the Localization of Viruses and Endosymbiotic Bacteria in Plant and Insect Tissues

荧光<em&gt;原位</em&gt;杂交（FISH）技术对病毒和植物和昆虫组织内共生细菌的本土化

Fluorescence in situ hybridization (FISH) is a name given to a variety of techniques commonly used for visualizing gene transcripts in eukaryotic cells ...

In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues

In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues

原位MHC-聚丙烯染色和定量分析确定抗原特异性 CD8 T 细胞在组织中的位置、丰度和表型

T cells are critical to many immunological processes, including detecting and eliminating virus-infected cells, preventing autoimmunity, assisting in ...

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 ...