Name: monitoring protein adsorption with solid-state nanopores
Uploaded: 2011-12-02T13:00:00
Description: Watch this Scientific Journal Video about Monitoring Protein Adsorption with Solid-state Nanopores at JoVE.com

作者想约翰Grazul（康奈尔大学），安德烈Marziali（在温哥华不列颠哥伦比亚大学）和外袍，科萨文森特（渥太华大学），以感谢他们的意见。这项工作是由美国国家科学基金会（DMR - 0706517和DMR - 1006332）和国立卫生研究院（R01 GM088403）资助部分经费。纳米孔钻在康奈尔大学材料研究中心（CCMR）从国家科学基金会支持的电子显微镜机制 - 材料研究科学和工程中心（MRSEC）程序（DMR 0520404）。氮化硅膜的制备是在康奈尔大学的纳米基金，国家纳米技术基础设施网络的成员，这是由国家科学基金会（格兰特ECS - 0335765）的支持。

1。氮化硅膜固态纳米孔的制造<ol><li>把费Tecnai F20的S /透射电镜一个200千伏的加速电压。如果使用不同的S /透射电子显微镜，加速电压应大于或等于200千伏9 </li><li> TEM样品架装入一个20纳米厚的SPI氮化硅窗口电网和清洁除去持有​​人的任何污染物与氧气30秒的等离子体。 </li><li>装入的S / TEM样品，并允许为真空抽空。一旦S /透射电镜抽空真空，发现在明场TEM模式通过寻找光明广场上的Ronc...

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自发到固态表面27-29上的蛋白质吸附在一些领域，如生物芯片的应用和设计一个新的功能混合生物材料类，是从根本上重要的。以往的研究表明，吸附到固体表面的蛋白质不显示横向调动或显著解吸率，因此蛋白质的吸附通常被认为是一个不可逆转的和非特异性的过程30-32。到固态表面的蛋白质吸附被认为是由于多种因素， 包括 13在固-液界面的蛋白质和活性基团的侧链之?...

<table border="1"><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> Tecnai F20的S /透射电子显微镜</td><td>费</td><td></td><td> S /透射电子显微镜需要加速电压≥200KV和场发射源。 </td></tr><tr><td> 20纳米厚的氮化硅膜窗口用于TEM </td><td> SPI </td><td> 4163SN - BA </td><td></td></tr><tr><td>轴突Axopatch 200B中膜片钳放大器</td><td>分子器件</td><td></td><td></td></tr><tr><td>轴突Digidata 1440A </td><td>分子器件</td><td></td><td></td></tr><tr><td> pCLAMP 10软件</td><td>分子器件</td><td></td><td>电数据采集和分析软件</td></tr><tr><td>硫酸</td><td> Fisher科学</td><td> A300 </td><td></td></tr><tr><td>过氧化氢</td><td> Fisher Scientific则</td><td> H325 </td><td></td></tr><tr><td>矽O型环</td><td>麦克马斯特 - 卡尔</td><td> 003 S70 </td><td>或者使用的PDMS </td></tr><tr><td>银线</td><td> Sigma - Aldrich公司</td><td> 348759 </td><td>对于电极</td></tr><tr><td> SPC技术，D SUB接触，针</td><td>纽瓦克</td><td> 9K4978 </td><td>对于电极</td></tr><tr><td>氯化钾</td><td>西格玛</td><td> P9541 </td><td></td></tr><tr><td>磷酸钾二元</td><td>西格玛</td><td> P2222 </td><td></td></tr><tr><td>磷酸二氢钾</td><td>西格玛</td><td> P5379 </td><td></td></tr><tr><td>硅橡胶</td><td>道康宁</td><td></td><td> Sylgar 184 Ëlastomer设置。制作室。 </td></tr><tr><td>佳柏，演员密封胶</td><td>世界精密仪器</td><td> KWIK - CAST </td><td>快速代理硅酮密封胶</td></tr><tr><td>热板</td><td> Fisher Scientific则</td><td></td><td></td></tr><tr><td>法拉第笼</td><td></td><td></td><td></td></tr></table>

monitoring protein adsorption with solid-state nanopores

固态纳米孔已被用来执行在单分子水平测量，以考察当地的结构和灵活性 1-6核酸， 蛋白质 7开展，8种不同的配体结合的亲和力。通过这些孔洞耦合9-12电阻脉冲技术，这样的测量，可以做各种各样的条件下，没有标签 3 。电阻脉冲技术，离子的盐溶液中引入纳米孔的两侧。因此，离子驱动从一个会议厅一侧的其他应用的跨膜电位，造成在一个稳定的电流。分析物的纳米孔的分割，导致在当前的定义良好的偏转，它可以分析提取单分子信息。使用这种技术，纳米孔壁的单个蛋白质的吸附作用，可以广泛的监视下条件13。蛋白质吸附的重要性越来越大，因为微流体器件尺寸的缩小，这些系统与单一蛋白质的相互作用成​​为一个令人关注的问题。这一协议描述为氮化物薄膜，它可以很容易地扩展到其他薄膜进行纳米孔钻，或功能化氮化物表面蛋白结合的快速检测。下一个广泛的解决方案和变性条件下，可以探索多种蛋白质。此外，该协议可用于探索使用纳米孔光谱的基本问题。

Watch this Scientific Journal Video about Monitoring Protein Adsorption with Solid-state Nanopores at JoVE.com

Monitoring Protein Adsorption with Solid-state Nanopores (Video) | JoVE

一个方法是使用固态纳米孔监测到无机表面蛋白质的非特异性吸附描述。该方法采用电阻脉冲的原则，允许实时和单分子水平探讨吸附。因为单一的蛋白质吸附过程是远离平衡，我们建议平行合成纳米孔阵列的就业，使表观一级反应速率常数蛋白质吸附定量测定以及和Langmuir吸附常数。

监测与固态纳米孔蛋白质吸附

Solid-state nanopores have been used to perform measurements at the single-molecule level to examine the local structure and flexibility of nucleic acids ...

Research

JoVE Journal

Bioengineering

Monitoring Protein Adsorption with Solid-state Nanopores

Registered Bioimaging of Nanomaterials for Diagnostic and Therapeutic Monitoring 

Registered Bioimaging of Nanomaterials for Diagnostic and Therapeutic Monitoring (Video) | JoVE

注册生物成像，纳米材料可用于诊断和治疗监测

Nanomedications can be carried by blood borne monocyte-macrophages into the reticuloendothelial system (RES; spleen, liver, lymph nodes) and to end ...

科研

生物工程

Fluorescence detection methods for microfluidic droplet platforms

Fluorescence detection methods for microfluidic droplet platforms (Video) | JoVE

荧光检测方法，为微流体液滴平台

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

Monitoring the Wall Mechanics During Stent Deployment in a Vessel

Monitoring the Wall Mechanics During Stent Deployment in a Vessel (Video) | JoVE

在一个容器的支架部署在监测的墙力学

Clinical trials have reported different restenosis rates for various stent designs1. It is speculated that stent-induced strain concentrations on the ...

Mass Cytometry: Protocol for Daily Tuning and Running Cell Samples on a CyTOF Mass Cytometer

Mass Cytometry: Protocol for Daily Tuning and Running Cell Samples on a CyTOF Mass Cytometer (Video) | JoVE

质量流式细胞术：协议，每日调整和执行细胞的样品上一个CyTOF的质量流式细胞仪

In recent years, the rapid analysis of single cells has commonly been performed using flow cytometry and fluorescently-labeled antibodies. However, the ...

Multi-analyte Biochip (MAB) Based on All-solid-state Ion-selective Electrodes (ASSISE) for Physiological Research

Multi-analyte Biochip MAB Based on All-solid-state Ion-selective Electrodes ASSISE for Physiological Research (Video) | JoVE

多分析物生物芯片（MAB），基于全固态离子选择性电极（ASSISE）的生理研究

Lab-on-a-chip (LOC) applications in environmental, biomedical, agricultural, biological, and spaceflight research require an ion-selective electrode (ISE) ...

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo (Video) | JoVE

多尺度孔梯度，聚电解质和间接监测改性金属植入物<em&gt;在体内</em

Metallic implants, especially titanium implants, are widely used in  clinical applications. Tissue in-growth and integration to these implants in  the ...

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests (Video) | JoVE

用于动态生长测试的温度，光照和pH监测的台式藻类光合生物反应器的构建和设置

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic ...

In Vesiculo Synthesis of Peptide Membrane Precursors for Autonomous Vesicle Growth

In Vesiculo Synthesis of Peptide Membrane Precursors for Autonomous Vesicle Growth (Video) | JoVE

在肽膜前体的维西图洛合成中,用于自主囊泡生长

Compartmentalization of biochemical reactions is a central aspect of synthetic cells. For this purpose, peptide-based reaction compartments serve as an ...

Functional Transcranial Doppler Ultrasound for Monitoring Cerebral Blood Flow

Functional Transcranial Doppler Ultrasound for Monitoring Cerebral Blood Flow (Video) | JoVE

功能颅内多普勒超声监测脑血流

Functional transcranial Doppler ultrasound (fTCD) is the use of transcranial Doppler ultrasound (TCD) to study neural activation occurring during stimuli ...

Labeling of Extracellular Vesicles for Monitoring Migration and Uptake in Cartilage Explants

Labeling of Extracellular Vesicles for Monitoring Migration and Uptake in Cartilage Explants (Video) | JoVE

标记细胞外囊泡以监测软骨外植体的迁移和摄取

Extracellular vesicles (EVs) are used in different studies to prove their potential as a cell-free treatment due to their cargo derived from their ...