Name: multimodal analytical platform on a multiplexed surface plasmon resonance imaging chip for the analysis of extracellular vesicle subsets
Uploaded: 2023-03-17T13:35:00
Description: Watch this Scientific Journal Video about Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets at JoVE.com

来自IVETh核心设施（巴黎）的Kelly Aubertin和Fabien Picot因拉曼成像实验而获得认可。Thierry Burnouf（台湾台北医科大学）和Zuzana Krupova（来自法国Helincourt）分别提供来自血小板和牛奶样本的EV样本。这项工作得到了勃艮第地区弗朗什-孔泰地区和欧洲 EIPHI 研究生院的支持（新手项目，2021-2024 年）。这项工作的一部分是使用CLIPP平台和FEMTO-ENGINEERING的RENATECH洁净室设施完成的，为此我们感谢Rabah Zeggari。

1. 金基板制备
注意：在金芯片上产生三种类型的表面：1）裸露表面，2）化学功能化和3）生物功能化（配体嫁接在C11C16层上）。从这一点开始，它们将分别被称为“裸”，“C11C16”和“配体”。
<ol><li>金基板制备： 注意：对于此协议，金生物芯片是在洁净室内部制造的。自制生物芯片由玻璃载玻片（SF11）组成，上面涂?...

<ol>
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qualification and quantification of platelet-derived microparticles.</a> Biosensors and Bioelectronics. 93, 250-259 (2017).</li><li>Ridolfi, 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=AFM-based+high-throughput+nanomechanical+screening+of+single+extracellular+vesicles.">AFM-based high-throughput nanomechanical screening of single extracellular vesicles.</a> Analytical Chemistry. 92 (15), 10274-10282 (2020).</li><li>Vorselen, 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+fluid+membrane+determines+mechanics+of+erythrocyte+extracellular+vesicles+and+is+softened+in+hereditary+spherocytosis.">The fluid membrane determines mechanics of erythrocyte extracellular vesicles and is softened in hereditary spherocytosis.</a> Nature Communications. 9 (1), 4960 (2018).</li><li>Hardij, 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=Characterisation+of+tissue+factor+bearing+extracellular+vesicles+with+AFM:+Comparison+of+air-tapping-mode+AFM+and+liquid+Peak+Force+AFM.">Characterisation of tissue factor bearing extracellular vesicles with AFM: Comparison of air-tapping-mode AFM and liquid Peak Force AFM.</a> Journal of Extracellular Vesicles. 2, 21045 (2013).</li><li>Jorgensen, 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=Extracellular+Vesicle+(EV)+Array:+Microarray+capturing+of+exosomes+and+other+extracellular+vesicles+for+multiplexed+phenotyping.">Extracellular Vesicle (EV) Array: Microarray capturing of exosomes and other extracellular vesicles for multiplexed phenotyping.</a> Journal of Extracellular Vesicles. 2, 20920 (2013).</li><li>Remy-Martin, 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=Surface+plasmon+resonance+imaging+in+arrays+coupled+with+mass+spectrometry+(SUPRA-MS):+Proof+of+concept+of+on-chip+characterization+of+a+potential+breast+cancer+marker+in+human+plasma.">Surface plasmon resonance imaging in arrays coupled with mass spectrometry (SUPRA-MS): Proof of concept of on-chip characterization of a potential breast cancer marker in human plasma.</a> Analytical and Bioanalytical Chemistry. 404 (2), 423-432 (2012).</li><li>Czamara, 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=Raman+spectroscopy+of+lipids:+A+review.">Raman spectroscopy of lipids: A review.</a> Journal of Raman Spectroscopy. 46 (1), 4-20 (2015).</li><li>Penders, 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=Single+particle+automated+Raman+trapping+analysis+of+breast+cancer+cell-derived+extracellular+vesicles+as+cancer+biomarkers.">Single particle automated Raman trapping analysis of breast cancer cell-derived extracellular vesicles as cancer biomarkers.</a> ACS Nano. 15 (11), 18192-18205 (2021).</li><li>Baek, S. J., Park, A., Ahn, Y. J., Choo, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Baseline+correction+using+asymmetrically+reweighted+penalized+least+squares+smoothing.">Baseline correction using asymmetrically reweighted penalized least squares smoothing.</a> Analyst. 140 (1), 250-257 (2015).</li><li>Daaboul, 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=Digital+detection+of+exosomes+by+interferometric+imaging.">Digital detection of exosomes by interferometric imaging.</a> Scientific Reports. 6, 37246 (2016).</li><li>Ertsgaard, C. 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最近使用最广泛的 EV 鉴定方法是蛋白质特异性免疫印迹法，用于确认 EV 的来源，TEM 以确认其结构，以及 NTA 以量化其在样品体积中的数量和大小分布3。然而，（生物）医学研究中对电动汽车的高度兴趣以及现有分析工具的局限性促使科学界开发用于电动汽车表征、区分和量化的新方法。
大多数开发侧重于将免疫标记或免疫捕获原理与用于EV检测的先进物理...

对诊断和治疗学1，2，3，4，5中的EV研究的兴趣日益浓厚，加上该领域面临的挑战，导致开发和实施各种方法来量化或表征这些囊泡的方法和技术。最广泛使用的EV鉴定方法是蛋白质特异性免疫印迹和蛋白质组学，以确认EV的来源，透射电子显微镜（TEM）以确认其结构，以及纳米颗粒跟踪分析（NTA）以量化其在样品体积中的数量和尺寸分布。
然而，这些技术本身都不能提供表征EV子集所需的所有信息。由于电动汽车生化和物理特性的多样性，其固有的异质性阻碍了可靠和可重复的全球分析，特别是对于混合物（粗样品）中所含的电动汽车。因此，电动汽车需要检测和表征方法，无论是单独还是一般，以补充其他更快但非选择性的方法6。
通过TEM（或cryoTEM）或AFM进行高分辨率成像，可以确定纳米分辨率为7，8，9，10，11，12的EV的形态和计量。然而，将电子显微镜用于生物物体（例如EV）的主要限制是需要真空来进行研究，这需要对样品进行固定和脱水。这种制备使得很难从观察到的结构转化为溶液内的EV形态。为了避免样品脱水，cryoTEM技术最适合EV表征13。它广泛用于确定电动汽车的超微结构。通过生物功能化的金纳米颗粒对囊泡进行免疫标记，还可以识别EV的特定亚群，并将其与复杂生物样品中存在的其他颗粒区分开来。然而，由于通过电子显微镜分析的EV数量很少，因此通常难以进行代表复杂和异质样品的表征。
为了揭示这种大小异质性，国际细胞外囊泡学会（ISEV）建议分析足够数量的宽场图像，并伴有较小的图像，以揭示具有高分辨率14的单个EV。AFM是用于研究电动汽车的光学方法和电子衍射技术的替代方案。该技术使用由柔性悬臂固定的锋利尖端，该悬臂逐行扫描沉积在一个支架上的样品，并通过反馈回路调整尖端与存在的元件之间的距离。这使得表征样品的形貌并收集形态力学信息成为可能15，16，17，18。EV可以在沉积在原子平坦的基板上后，或者在由抗体，肽或适配体功能化的特定底物上捕获以表征各种亚群后通过AFM进行扫描18，19。由于AFM能够在复杂的生物样品中定量并同时探测EV的结构、生物力学和膜生物分子含量，而无需预处理、标记或脱水，因此现在越来越多地用于在温度和介质的生理条件下以精细和多参数的方式表征EV。
本文提出了一种使用核心金生物芯片的方法，该方法能够以多重形式进行（生物）化学功能化。该基板是强大的分析平台的基石，该平台结合了通过表面等离子体共振对EV亚群进行生物检测，一旦EV被吸附/接枝或免疫捕获到芯片上，AFM就可以对EV进行计量和形态力学表征。结合芯片上捕获的EV子集的拉曼特征，该分析平台能够以无标记的方式鉴定生物样品中存在的EV，而无需任何分析前步骤。本文表明，强大的技术与高度严格的底物制备和数据采集方法相结合，使EV分析具有深入性、确定性和稳健性。
该方法的原理是制备金基底，吸附/嫁接或捕获EV亚型，并通过AFM扫描它们以估计每个EV亚群的大小和形态。此外，这些吸附的电动汽车通过拉曼光谱进行分析。事实上，这种底物可以呈现三种日益复杂的界面：裸界面、化学功能化界面或配体微阵列。在描述协议的不同步骤之前，读者参考 图1中纳米生物分析平台（NBA）方法的示意图，该方法结合了表面等离子体共振成像（SPRi），AFM和光谱学。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64210/64210fig01v2.jpg"> 图1：纳米生物分析平台。该方法结合了（A）表面等离子体共振成像，（B）原子力显微镜和红外/拉曼（纳米）光谱，所有这些都在同一基板上 - 多路复用金芯片。缩写：NBA = 纳米生物分析平台;SPRi = 表面等离子体共振成像;AFM = 原子力显微镜;EV = 细胞外囊泡。 <a href="https://www.jove.com/files/ftp_upload/64210/64210fig01largev2.jpg" target="_blank">请点击此处查看此图的大图。</a>
核心金生物芯片构成了该平台的核心，因为所有无标记表征技术都是在该生物芯片上进行的。根据 EV 表征的需求（全球/总 EV 或 EV 子集）和所用方法的局限性/要求，已经开发了三种类型的金生物芯片表面：“裸”，化学功能化的“C11/C16”或配体生物功能化的，称为“配体”金表面。
这种被称为“裸体”的裸生物芯片能够简单地将电动汽车吸附在黄金上。可以选择使用的缓冲液，并以被动方式（孵育然后冲洗步骤）或在流动下（在SPRi中）监测其实现这种吸附。此外，这种被动吸附既可以在整个芯片上（作为宏阵列）实现，也可以使用微量移液器点样器定位在微阵列中。“流动下程序”允许研究人员跟踪动力学和EV吸附水平。当化学层界面可能干扰分析方法（例如，用于拉曼光谱）时，在裸金衬底上采用这种方法。
 化学功能化的生物芯片称为“C11/C16”，用于通过与硫醇酸盐形成伯酰胺键，在金表面上共价结合的EV的致密而坚固的“地毯”，目的是获得EV样品的全局视图。事实上，在这种情况下，金被巯基-1-十一醇（11-MUOH：“C11”）和巯基-1-十六烷酸（16-MHA：“C16”）的硫代酸盐混合物官能化，并且一小部分硫醇酸盐被化学活化以建立与靶标的共价结合。同样，这种策略可以被动地（孵育然后冲洗步骤，无论是在“宏阵列”中还是在多个微阵列中使用微量移液器点样器）或在流速下（在SPRi中）实现，以跟踪EV接枝在金表面上的动力学和水平。
配体 生物功能化的生物芯片，称为 “配体”，被化学激活以共价移植不同的配体（例如，抗体，受体），以选择性地捕获（亲和力）在生物样品中共存的不同EV亚群。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CD41a antibody</td>
			<td>Diaclone SAS (France)</td>
			<td>447528</td>
			<td></td>
		</tr>
		<tr>
			<td>CP920</td>
			<td>Microparticles GmbH, Germany</td>
			<td>448303</td>
			<td></td>
		</tr>
		<tr>
			<td>DXR3xi&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>T1502</td>
			<td></td>
		</tr>
		<tr>
			<td>EDC</td>
			<td>Sigma</td>
			<td>A6272</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanolamine</td>
			<td>Sigma</td>
			<td>P5368-10PAK</td>
			<td></td>
		</tr>
		<tr>
			<td>Evs derived from platelet concentrates</td>
			<td>Collaboration : Pr T. Burnouf (TMU, Taipei)</td>
			<td>S2889</td>
			<td></td>
		</tr>
		<tr>
			<td>Evs from bovine milk</td>
			<td>Collaboration : Dr Z. Krupova (Excilone, Helincourt - France)</td>
			<td>3450</td>
			<td></td>
		</tr>
		<tr>
			<td>Glutaraldehyde</td>
			<td>Sigma</td>
			<td>56845</td>
			<td></td>
		</tr>
		<tr>
			<td>Gwyddion&#160;</td>
			<td></td>
			<td>853.223.020</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetron sputtering</td>
			<td>PLASSYS</td>
			<td>SAB5300165</td>
			<td></td>
		</tr>
		<tr>
			<td>mercapto-1-hexadecanoic acid</td>
			<td>Sigma</td>
			<td>G5882</td>
			<td></td>
		</tr>
		<tr>
			<td>Mercapto-1-undecanol&#160;</td>
			<td>Sigma</td>
			<td>O8001</td>
			<td></td>
		</tr>
		<tr>
			<td>Mountains SPIP ones</td>
			<td>Digital Surf</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NanoWizard 3 Bioscience&#160;</td>
			<td>Bruker-JPK&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Octyl Glucoside (OG)&#160;</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ovalbumine antibody</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffer Saline (PBS)</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rat Albumin Serum (RSA)</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium acetate buffer&#160;</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SPR-Biacore 3000</td>
			<td>GE Healthcare/ Cytiva life sciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SPRi Biochip</td>
			<td>MIMENTO technology platform</td>
			<td></td>
			<td>The biochips were produced in-house in the clean room, Besancon</td>
		</tr>
		<tr>
			<td>SPRi Plex II</td>
			<td>Horiba Scientific&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sulfo-NHS</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

multimodal analytical platform on a multiplexed surface plasmon resonance imaging chip for the analysis of extracellular vesicle subsets

细胞外囊泡（EV）是由所有细胞产生的膜衍生的微小囊泡，直径范围为50至数百纳米，用作细胞间通讯的手段。它们正在成为各种疾病的有前途的诊断和治疗工具。细胞使用两种主要的生物发生过程来生产大小、组成和含量不同的 EV。由于它们的大小、组成和细胞来源高度复杂，它们的表征需要结合分析技术。该项目涉及开发新一代多参数分析平台，具有更高的吞吐量，用于表征电动汽车的亚群。为了实现这一目标，这项工作从该小组建立的纳米生物分析平台（NBA）开始，该平台允许基于多重生物传感方法与原子力显微镜（AFM）的计量和形态力学分析相结合的EV进行原始研究微阵列生物芯片上捕获的囊泡靶标。目的是通过拉曼光谱的表型和分子分析来完成这项 EV 研究。这些发展使得能够提出一种多模态且易于使用的分析解决方案，用于区分具有临床潜力的生物体液中的EV亚群

确定配体接枝的最佳pH条件 用于制备生物芯片的不同配体根据pH值及其与硫代酸盐化学层相互作用的可用性进行测试（图3）。将配体稀释在不同的pH值的乙酸盐缓冲液中，并注入用C11C16层化学官能化的生物芯片上。将溶液随机注入表面上，并在每次配体注射后注射去垢剂（40mM的OG）以恢复基线。该 预浓缩 测试可以确定每个配体移植的最佳pH值。在...

Watch this Scientific Journal Video about Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets at JoVE.com

Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets

本文提出了新一代多参数分析平台，具有更高的通量，用于表征细胞外囊泡亚群。该方法基于多重生物传感方法与原子力显微镜的计量和形态力学分析相结合，结合拉曼光谱，以鉴定捕获在微阵列生物芯片上的囊泡靶标。

多路复用表面等离子体共振成像芯片上的多模态分析平台，用于分析细胞外囊泡亚群

Extracellular vesicles (EVs) are membrane-derived, tiny vesicles produced by all cells that range from 50 to several hundreds of nanometers in diameter ...

EV的复杂性使得很难识别感兴趣的亚群，并且该协议可以通过其表型，尺寸特征和纳米力学特性来检测它们。这种技术组合是一种无标记方法，它使我们能够从条件培养基和血浆中实时检测电动汽车。该技术能够表征感兴趣的纳米颗粒的深层子集，以便用作病理学的生物指示剂，或也用作药物递送的纳米颗粒。由于这是一种非常灵敏的方法，因此在注入感兴趣的样品之前必须注意生物芯片的制备。在对芯片进行包衣和功能化后，将生物芯片在 200 毫摩尔乙基二甲氨基丙基碳化二亚胺/N-羟基琥珀酰亚胺和 50 毫摩尔/升 N-羟基琥珀酰亚胺的混合物中孵育至少 30 分钟在室温下黑暗中。使用观察剂，加入300纳升配体溶液，并将生物芯片在声浴下孵育30分钟。用超纯水从顶部清洗生物芯片。加入一滴与棱镜折射率相同的油滴，在生物芯片和棱镜之间形成均匀的薄层，并将其轻轻地放在与生物芯片折射率相同的棱镜上。对于表面等离子体共振成像，将生物芯片安装在SPRi系统上。导航到软件左侧的下拉菜单，然后单击工作目录。找到可见不同斑点的图像，然后单击以选择此图像。然后写下配体家族的名称，然后单击完成物种定义。将带有光标的黑线拖动到最佳工作角度。单击将镜像移动到工作角度，然后选择一个工作角度。现在点击动力学。当软件提示用户定义阴性对照时，此时选择无阴性对照。以每分钟50微升的速度注射大鼠血清白蛋白四分钟。以每分钟20微升的速度注射乙醇胺10分钟，以使表面上仍然存在并具有反应性的羧基失活。之后，通过以每分钟 50 微升的速度注射 40 毫摩尔 OG 来洗涤生物芯片，持续四分钟。在生物功能化芯片上以特定浓度注射细胞外囊泡，同时跟踪不同点上囊泡相互作用的动力学。还要确定反射率的值和相互作用的水平。一旦稳定下来，在芯片上注射戊二醛，在进行AFM成像之前将细胞外囊泡固定在它们所在的位置。将载玻片上面罩顶部的生物芯片对齐。使用AFM顶部的CCD相机将悬臂定位在必须扫描的正确位置。在接触模式下从三个到五个大区域到小区域开始AFM采集。接下来，通过首先选择高度通道，使用JPK数据处理软件处理AFM图像。选择要从每条线中减去的多项式拟合以获得拉直的扫描线。选择金晶粒上的高度阈值以消除表面上的粗糙度。晶粒提取模块使用 8.5 纳米的高度阈值标记晶粒。分析白蛋白钝化后的多重生物芯片。没有默认值的芯片。芯片由于斑点融合、接枝较弱或气泡或污染物而存在一些缺陷。并显示了没有微阵列的裸金芯片，用于检查细胞外囊泡在金上的吸附。在多重生物芯片上的捕获实验显示出不同配体的良好反射率信号，以及不同配体的良好信噪比，因为阴性对照的反应可以忽略不计。注射细胞外囊泡样品后的细胞外囊泡吸附显示出高反射率信号，表明这些囊泡能够吸附并保持在金芯片上稳定。细胞外囊泡加载后，在生物芯片上生成细胞外囊泡的大规模和小规模AFM图像。高分辨率近距离观察可实现细胞外囊泡的计量。生物芯片上细胞外囊泡的有效直径范围为30至300纳米，其中绝大多数在60纳米左右。在空气和液体中获得的结果具有可比性。吸附在裸芯片上的细胞外囊泡的拉曼光谱显示出清晰的光谱，其峰主要对应于亚甲基振动，亚甲基振动与细胞外囊泡膜的脂质有关。严格准备您的生物芯片非常重要，以便能够以可重复的方式准确检测您的电动汽车。可以使用蛋白质印迹和纳米颗粒示踪分析等常规方法来关联和确认表型和尺寸模式。此外，多组学分析甚至有望更深入地研究 EV 分子表征和潜在的子集区分。

Research

JoVE Journal

Biology

Assembly, Loading, and Alignment of an Analytical Ultracentrifuge Sample Cell

大会，加载和分析超速离心机样品池对齐

The analytical ultracentrifuge (AUC) is a powerful biophysical tool that allows us to record macromolecular sedimentation profiles during high speed ...

科研

生物学

Lensless On-chip Imaging of Cells Provides a New Tool for High-throughput Cell-Biology and Medical Diagnostics

无透镜片上的细胞成像提供了一种高通量细胞生物学和医疗诊断的新工具

Conventional optical microscopes image cells by use of objective lenses that work together with other lenses and optical components. While quite ...

Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

采用芯片识别亚型特定的靶基因的全基因组分析

Recruitment of transcriptional and epigenetic factors to their targets is a key step in their regulation. Prominently featured in recruitment are the ...

Electron Spin Resonance Micro-imaging of Live Species for Oxygen Mapping

电子自旋共振氧测绘活体物种显微成像

This protocol describes an electron spin resonance (ESR) micro-imaging method for three-dimensional mapping of oxygen levels in the immediate environment ...

Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions

Avidity-based Extracellular Interaction Screening AVEXIS for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions

基于亲和力，外的相互作用筛选（AVEXIS）外低亲和力受体 - 配体相互作用的可扩展检测

Extracellular protein:protein interactions between secreted or membrane-tethered proteins are critical for both initiating intercellular communication and ...

gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair

gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair

基因组DNA富集转座为基础的技术的整个序列的农工商分析<em&gt; BRCA1基因</em&gt;<em&gt; BRCA2</em&gt;和9个基因参与DNA损伤修复

The widespread use of Next Generation Sequencing has opened up new avenues for cancer research and diagnosis. NGS will bring huge amounts of new data on ...

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

膜蛋白的实时测量：受体相互作用使用表面等离子体共振（SPR）

Protein-protein interactions are pivotal to most, if not all, physiological processes, and  understanding the nature of such interactions is a central ...

Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis

从富集组织及消化细胞外基质蛋白的成肽质谱分析

The extracellular matrix (ECM) is a complex meshwork of cross-linked proteins that provides biophysical and biochemical cues that are major regulators of ...

Measurement and Analysis of Extracellular Acid Production to Determine Glycolytic Rate

测量外产酸和分析，以确定糖酵解率

Extracellular measurement of oxygen consumption and acid production is a simple and powerful way to monitor rates of respiration and glycolysis1. Both ...

Nanoparticle Tracking Analysis for the Quantification and Size Determination of Extracellular Vesicles

细胞外囊泡的量化和尺寸测定纳米粒子跟踪分析

The physiological and pathophysiological roles of extracellular vesicles (EVs) have become increasingly recognized, making the EV field a quickly evolving ...