这项工作是联合太阳能计划（JSP）Hyet太阳能基金会手背上Fundamenteel的Onderzoe​​k der质料的FOM，这是荷兰科学研究组织（NWO）的一部分。

1。制备块切片<ol><li>治疗技术等级3“硅片在空气等离子清洁，持续30秒，然后将其暴露在（十三1，1,2,2 -四氢甲基）三氯硅烷的蒸气一小时。 注意：这一步是必要的前步骤1.4，以防止环氧树脂附着到硅晶片。 </li><li>沉积一层金（通常为100 nm厚的，它定义了导线的宽度）通过特氟隆主站（即定义得到的线的长度为0.5毫米，1毫米，或1.5毫米）到预处理过的硅晶片上。 </li><li>量在整个晶片〜8.5毫升环氧预聚物Epofix的和为3个小时，在60℃下固化</li><li>金层剥离模板，通过仔细地，黄金仍然连接到固化后的环氧从晶片剥离环氧。在硅晶片和环氧树脂之间的界面插入剃刀刀片的边缘，然后轻轻剥离环氧树脂层在硅晶片。由于黄金附着力差氟化硅晶片（步骤1.1）的黄金功能仍然坚持环氧树脂。 注意：要小心，不要破坏硅片，否则会损坏硅颗粒钻石刀切片步骤。 </li><li>相同的聚四氟乙烯掩模被放置在黄金功能，但横向偏移的〜80％的最短尺寸的金功能，通过特氟隆主沉积的铝或银层。该层的厚度将定义的纳米间隙的金线之间的间距。的下限取决于金属，但〜5nm的铝和银，低于该层成为不连续的。 注：这个偏置电压将最终定义两个金电极之间的重叠长度，你可以用微标尺测量。 </li><li> 5纳米以下差距：浸入环氧模板精简黄金1毫米的解决方案的alkanedithiol在乙醇（或任何溶剂，不溶胀环氧树脂）过夜，用氮气吹洗（减轻自发形成的二硫化物）在一个封闭的腔室。 （在本文中，我们使用1,12 - dodecanedithiol 1,14 tetradecanedithiol的或1,16 - hexadecanedithiol的的生产各种宽度小于3纳米的差距。）拆下的模板剥离金环氧树脂基板从SAM形成的解决方案。用乙醇冲洗，并用氮气干燥，在干燥之前，持续2分钟，在60。 </li><li>如果使用SAM，这里，回到聚四氟乙烯掩模的环氧基板上，但横向偏移〜80％的最短尺寸的金功能。如果使用金属，面膜已经是1.5步后的位置，所以不改变位置的面具后沉积的铝或银。 </li><li>沉积第二层的黄金或通过掩模的任何其它金属。此层通常可以由相同的金属上面的厚度，作为第一个（在这种情况下，100 nm厚）。 </li><li>消除特氟隆口罩，注意不要划伤的功能，这将导致破碎的纳米线。 </li><li>重新嵌入在Epofix预聚物（约8.5毫升），在整个基板，至少3个小时，在60固化。 </li><li>切出来的功能，使用一个珠宝商的锯（成〜4×10毫米件），并放置到一个单独的井在聚乙烯“棺材”切片模具。 </li><li>填充模具与Epofix预聚物，并在60℃下过夜固化</li></ol> 2。切片<ol><li>从聚乙烯模具中取出一块，将其安装在样品架。 </li><li>将修剪附件样品架，将其安装在超薄切片。 </li><li>剃刀刀片，用乙醇清洗以除去润滑剂和金属碎片下的超薄切片的体视镜检查刀片的边缘。任何剩余的碎片会损坏钻石刀切片过程中。修剪块的宽度的钻石刀（瓦特E使用2毫米或4的毫米Diatome超35°）呈梯形（因为它是最稳定的形状，切片）。一些ultramicrotomes 注：使用修剪附件安装切割臂，但我们取得更好的成绩与刀片。 </li><li>玻璃刀平行地面对块的底部边缘的边缘对齐。 </li><li>启动与配备面对的块定义一个光滑的表面上的玻璃刀的超薄切片机（我们使用的Leica EMUC-6）的预切割。 </li><li> ，制作金属结构，取代在1毫米/秒或50纳米的钻石刀，重新对齐，块部分为100nm的玻璃刀为0.6毫米/秒。 epofix部分是稳定的下降到约30 nm。 注：一个易于验证的部分的厚度是它的色彩，作为厚度的函数的变化可预测的和不依赖于已使用的树脂，该树脂可参考卡。 16 </li&gt; <li>收藏刀水库中的水的表面结构包含环氧部分，无论是单独使用一个完美的环路（电子显微学）或作为色带的几个部分的Si/SiO2（SEM）或SiO 2（电气测量）衬底上放置基板下表面的水，并慢慢提高。 </li><li>部分在60℃下干燥3小时，以提高其对基材的附着力。 </li><li>灰环氧树脂，使样品的氧等离子体（15分钟1毫巴足以消除所有痕迹的环氧100或50 nm厚部分）。 注：如果制造电气测量的纳米结构，这一步必须在步骤4之后进行的。 </li></ol> 3。牺牲层蚀刻<ol><li>铝：将在2M的HCl水溶液2小时的部分含有铝。白银：暴露部分氧等离子体10最小。 注：材料的选择可以是湿法蚀刻（用HCl）或干法蚀刻（使用氧等离子体），但是银可以通过湿蚀刻除去，以及。 </li><li>地空导弹：等离子体处理部分蚀刻地对空导弹，但我们已经在我们的努力到什么程度表征不成功。 </li></ol> 4。电气测量<ol><li>将环氧部分SiO2衬底上已彻底清洗（ 例如使用piraña溶液）和干燥（步骤2.7和2.8）。 </li><li>安装基板，光学显微镜下或超微立体镜。 </li><li>应用滴银膏（或碳墨），在每个部分上的电线的两端。这些嵌入的金属结构，将显示一条黑线（从金/环氧接口）或较厚的金的结构的情况下，（从沉积步骤），直接可见的。在这两种情况下，应适用​​滴足够远，从中心到短的纳米间隙。 </li></ol>

<ol>
	<li>Lipomi, D. J., Martinez, R. V., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+thin+sectioning+(nanoskiving)+to+fabricate+nanostructures+for+electronic+and+optical+applications.">Use of thin sectioning (nanoskiving) to fabricate nanostructures for electronic and optical applications.</a> Angew. Chem. Int. Ed. 50 (37), 8566-8583 (2011).</li><li>Xu, Q., Rioux, R. M., Dickey, M. D., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoskiving:+A+new+method+to+produce+arrays+of+nanostructures.">Nanoskiving: A new method to produce arrays of nanostructures.</a> Acc. Chem. Res. 41 (12), 1566-1577 (2008).</li><li>Xu, Q., Rioux, R. M., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+complex+metallic+nanostructures+by+nanoskiving.">Fabrication of complex metallic nanostructures by nanoskiving.</a> ACS Nano. 1 (3), 215-227 (2007).</li><li>Reed, M. A., Zhou, C., Muller, C. J., Burgin, T. P., Tour, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conductance+of+a+molecular+junction.">Conductance of a molecular junction.</a> Science. 278 (5336), 252-254 (1997).</li><li>Chen, W., Ahmed, H., Nakazoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coulomb+blockade+at+77+k+in+nanoscale+metallic+islands+in+a+lateral+nanostructure.">Coulomb blockade at 77 k in nanoscale metallic islands in a lateral nanostructure.</a> Appl. Phys. Let. 66 (24), 3383-3384 (1995).</li><li>Morpurgo, A. F., Marcus, C. M., Robinson, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+fabrication+of+metallic+electrodes+with+atomic+separation.">Controlled fabrication of metallic electrodes with atomic separation.</a> Appl. Phys. Let. 74 (14), 2084-2086 (1999).</li><li>Paska, Y., Haick, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+cross-linking+changes+within+a+self-assembled+monolayer+in+a+nanogap+junction:+A+tool+for+investigating+the+intermolecular+electronic+coupling.">Systematic cross-linking changes within a self-assembled monolayer in a nanogap junction: A tool for investigating the intermolecular electronic coupling.</a> J. Am. Chem. Soc. 132 (6), 1774-1775 (2010).</li><li>Park, J., Pasupathy, A. N., Goldsmith, J. I., Chang, C., Yaish, Y., Petta, J. R., Rinkoski, M., Sethna, J. P., Abruna, H. D., McEuen, P. L., Ralph, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coulomb+blockade+and+the+kondo+effect+in+single-atom+transistors.">Coulomb blockade and the kondo effect in single-atom transistors.</a> Nature. 417 (6890), 722-725 (2002).</li><li>Nagase, T., Kubota, T., Mashiko, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+nano-gap+electrodes+for+measuring+electrical+properties+of+organic+molecules+using+a+focused+ion+beam.">Fabrication of nano-gap electrodes for measuring electrical properties of organic molecules using a focused ion beam.</a> Thin Solid Films. 438-439, 374-377 (2003).</li><li>Kubatkin, S., Danilov, A., Hjort, M., Cornil, J., Br&#233;das, J. -. L., Stuhr-Hansen, N., Hedeg&#229;rd, P., Bj&#248;rnholm, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-Electron+Transistor+of+a+Single+Organic+Molecule+with+Access+to+Several+Redox+States.">Single-Electron Transistor of a Single Organic Molecule with Access to Several Redox States.</a> Nature. 425 (6959), 698-701 (2003).</li><li>Notargiacomo, A., Foglietti, V., Cianci, E., Capellini, G., Adami, M., Faraci, P., Evangelisti, F., Nicolini, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atomic+force+microscopy+lithography+as+a+nanodevice+development+technique.">Atomic force microscopy lithography as a nanodevice development technique.</a> Nanotechnology. 10 (4), 458-463 (1999).</li><li>Qin, L., Park, S., Huang, L., Mirkin, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On-wire+lithography.">On-wire lithography.</a> Science. 309 (5731), 113-115 (2005).</li><li>Hatzor, A., Weiss, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+rulers+for+scaling+down+nanostructures.">Molecular rulers for scaling down nanostructures.</a> Science. 291 (5506), 1019-1020 (2001).</li><li>Pourhossein, P., Chiechi, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directly+addressable+sub-3+nm+gold+nano-gaps+fabricated+by+nanoskiving+using+self-assembled+monolayers+as+templates.">Directly addressable sub-3 nm gold nano-gaps fabricated by nanoskiving using self-assembled monolayers as templates.</a> ACS Nano. 6, 5566-5573 (2012).</li><li>Lipomi, D. J., Ilievski, F., Wiley, B. J., Deotare, P. B., Lon&#269;ar, M., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+fabrication+and+magnetic+positioning+of+metallic+and+polymeric+nanowires+embedded+in+thin+epoxy+slabs.">Integrated fabrication and magnetic positioning of metallic and polymeric nanowires embedded in thin epoxy slabs.</a> ACS Nano. 3 (10), 3315-3325 (2009).</li><li>Mays, R. L., Pourhossein, P., Savithri, D., Genzer, J., Chiechi, R. C., Dickey, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thiol-containing+polymeric+embedding+materials+for+nanoskiving.">Thiol-containing polymeric embedding materials for nanoskiving.</a> Journal of Materials Chemistry C. , (2013).</li><li>Thuo, M. M., Reus, W. F., Nijhuis, C. A., Barber, J. R., Kim, C., Schulz, M. D., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odd-even+effects+in+charge+transport+across+self-assembled+monolayers.">Odd-even effects in charge transport across self-assembled monolayers.</a> J. Am. Chem. Soc. 133 (9), 2962-2975 (2011).</li><li>Song, H., Kim, Y., Jeong, H., Reed, M. A., Lee, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+Tunneling+Transport+in+Molecular+Junctions.">Coherent Tunneling Transport in Molecular Junctions.</a> J. Phys. Chem. C. 114 (48), 20431-20435 (2010).</li><li>Wang, W. Y., Lee, T., Reed, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+Electron+Conduction+in+Self-Assembled+Alkanethiol+Monolayer+Devices.">Mechanism of Electron Conduction in Self-Assembled Alkanethiol Monolayer Devices.</a> Phys. Rev. B. 68 (3), 035416 (2003).</li><li>Weiss, E. A., Chiechi, R. C., Kaufman, G. K., Kriebel, J. K., Li, Z., Duati, M., Rampi, M. A., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+defects+on+the+electrical+characteristics+of+Mercury-Drop+junctions:+Self-Assembled+monolayers+of+n-+Alkanethiolates+on+rough+and+smooth.">Influence of defects on the electrical characteristics of Mercury-Drop junctions: Self-Assembled monolayers of n- Alkanethiolates on rough and smooth.</a> 129 (14), 4336-4349 (2007).</li></ol>

没有利益冲突的声明。

在本文中，我们展示的制造使用nanoskiving纳隙结构。这个实验简单的方法，使生产的纳米结构，在一秒钟的速度，所有三个维度的控制权。的差距大小是由铝和银的结合要么牺牲层或二硫醇（它提供了一个决议作为一个小）自组装单分子膜。纳米结构可被定位在任意衬底上的手，和它们直接电寻址，这是一个唯一的属性nanoskiving的。这项技术也产生高度均匀的结构，但是，敏感的振​​动，从而改...

本文详细介绍了制造电寻址，高宽比黄金纳米线使用单纳米真空间隙分离沉积铝和银作为牺牲隔离层的空白&gt; 5 nm和自组装单分子膜（SAMS）alkanedithiols小的差距为1.7 nm。我们制作了这些纳米结构没有一个干净的房间或切片边缘光刻称为nanoskiving的一种形式，使用超薄切片机，牺牲间隔物隔开黄金夹层结构的任何光刻工艺。1-3此方法的组合沉积薄金属电影和使用超薄切片机切片。主升压在nanoskiving的切片超薄切片，用超薄切片机配备有钻石刀，其安装在装满水的产生是〜30nm的薄板坯的小船。广泛用于ultramicrotomes薄的样品用于制备具有光学成像，或选择罗恩显微镜和许多最有经验的从业者来自生物或医学背景的超薄切片。有几种方法包括机械破发路口，4电子束光刻5，电化学电镀，6，7电迁移，聚焦离子束光刻，9影子蒸发，10扫描探针和原子力显微镜，11线光刻制造nanogaps ，12和13分子的统治者。所有这些方法都有自己的特点和应用，但生产和寻址nanogaps有用的电话号码，并精确控制尺寸的差距仍然是一个挑战。此外，这些方法具有运行费用高，它们仅限于类的材料，可以生存的蚀刻工艺，在分辨率是有限的。 Nanoskiving使的快速制造电寻址纳米线的spacinGS的单纳米板凳上顶。我们感兴趣的是快速原型纳米结构的分子电子学，纳米制造电极技术不需要专门或耗时14块一次，它可以产生数百成千上万的纳米结构（连续）的需求。然而，这项技术不仅限于自组装膜或分子电子学，是一种通用的方法制备两种纳米结构之间的间隙。在本文中我们使用银，铝，和自组装膜作为牺牲层，以产生不同大小的金纳米线之间的间隙，但该方法并不限于这些材料（或金属纳米线）。电线挑地方，带磁性对齐兼容，因此，他们可以放在任意基板的nanoskiving 15的另一个优势是，它可以提供所有三个维度的控制权。在基板的地形（X），由样品的尺寸沉积膜（Y）和生产的超薄切片机（Z）的板坯的厚度的厚度。 图1总结了用于生产纳米线与所定义的间隔的步骤。黄金功能（1-2毫米长）通过蒸发沉积，通过聚四氟乙烯掩模到硅基片上。 （电子显微镜epofix版）环氧预聚物倒入覆盖金时，环氧树脂固化后，环氧树脂被从晶片分离（ 即通过模板剥离）的功能，在整个晶片，黄金功能保持粘附到环氧。对于金属牺牲层，铝或银的蒸发所需厚度通过聚四氟乙烯掩模的一个偏移量为200 - 500微米以上的金功能。要分5纳米的差距，形成一个SAM淹没黄金的功能，在适当的硫醇过夜1毫米的乙醇溶液。通过放置在特氟隆荫罩淀积金（或其它金属）的第二组金功能（覆盖在银，铝或SAM）的第一层，其偏移量为200 - 500μm的相对于第一蒸发。此偏移量将最终确定的最长尺寸的间隙，并用微型嵌入环氧树脂中的切片的整个结构的前标尺，它可以精确地测量。然后，整个结构被嵌入在一个块中的环氧树脂，然后用超薄切片机切片可能准备好。采样臂持有制备的块作为钻石刀朝它的进步，在控制步骤，将板坯的厚度定义。将得到的部分浮在水面上的小船。

<table border="1">
<tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent/Material</td>
 </tr>
 <tr>
 <td>Epofix epoxy resin </td>
 <td> Electron Microscopy </td>
 <td> 1232 </td>
 <td> </td>
 </tr>
 <tr>
 <td></td>
 <td> Sciences </td>
 <td> </td>
 <td> </td>
 </tr>
 <tr>
 <td>Gold </td>
 <td> Schone Edelmetaal B.V</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Aluminum </td>
 <td> Umicore Materials AG </td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Silver </td>
 <td> Umicore Materials AG </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td>(tridecafluoro-1,1,2,2, </td>
 <td> ABCR GmbH co.KG</td>
 <td> 78560-45-9 </td>
 <td></td>
 </tr>
 <tr>
 <td>-tetrahydrooctyl) </td>
 <td> </td>
 <td> </td>
 <td> </td>
 </tr>
 <tr>
 <td>trichlorosilane </td>
 <td></td>
 <td> </td>
 <td> </td>
 </tr>
 <tr>
 <td>,12-dodecanedithiol </td>
 <td> Home-synthesised </td>
 <td> </td>
 <td>According to: Akkerman et. al., Nature. 441, 69-72 (2006)</td>
 </tr>
 <tr>
 <td>,14-tetradecanedithiol </td>
 <td> synthesized in house </td>
 <td> </td>
 <td> According to: Akkerman et. al., Nature. 441, 69-72 (2006)</td>
 </tr>
 <tr>
 <td>,16-hexadecanedithiol </td>
 <td> synthesized in house </td>
 <td> </td>
 <td>According to: Akkerman et. al., Nature. 441, 69-72 (2006)</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Thermal deposition system </td>
 <td> home-built </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td>Ultramicrotome </td>
 <td> Leica Microsystems </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td>Dimanod knife ultra 35 </td>
 <td> Diatome </td>
 <td> DU3540</td>
 <td></td>
 </tr>
 <tr>
 <td>Dimanod knife ultra 45 </td>
 <td> Scimed GMBH </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td>Scanning electron microscope </td>
 <td> JOEL </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td>Source meter </td>
 <td> Keithley </td>
 <td> </td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Table 1. Tables of Specific Reagents and Equipment.</td>
 </tr>
 </tbody>
</table>

fabricating nanogaps by nanoskiving

有几种方法制造与控制间距nanogaps，但在两个电极之间，并产生在实际数量亚纳米间距的精确的控制仍然是具有挑战性的。 ，使用nanoskiving，这是一种形式的边缘光刻，纳隙电极的制备是一个快速，简单和强大的技术。这种方法是完全是机械的过程，其中不包括任何照片或电子束光刻步骤，并且不需要任何特殊的设备或基础设施，例如无尘室。 Nanoskiving用于制造电寻址nanogaps与所有三个维度的控制权，这些结构定义的最小尺寸牺牲层（Al或Ag）或自组装单分子膜的厚度。这些电线可以被手动定位在运输上的水珠，并直接电寻址，没有进一步的光刻技术需要将它们连接到一个静电。

我们准备将两个金属牺牲层作为间隔：铝和银纳隙结构。我们铭刻这些层获得所需的厚度差距。 “协议”一节中所描述的，切片后暴露结构含银的氧等离子体，和那些含有铝HCl水溶液， 图2所示的扫描电子显微照片（中小型企业），所得到的纳米线，纳米级的分离。在这两种情况下，差距清晰可见，直接测量。为了获得小于3纳米的差距，我们采用自组装单分子层1,12 - dodecanedithiol的?...

Watch this Scientific Journal Video about Fabricating Nanogaps by Nanoskiving at JoVE.com

Fabricating Nanogaps by Nanoskiving

电寻址，高纵横比（&gt; 1000:1）制造金属纳米线的差距可以使用铝和银的牺牲层自组装单分子膜为模板的单纳米分离。这些纳隙结构没有一个干净的房间或任何边缘光刻称为nanoskiving的的一种形式由光或电子束光刻工艺制作。

制造由Nanoskiving Nanogaps的

There are several methods of fabricating nanogaps with controlled spacings, but the precise control over the sub-nanometer spacing between two ...

Research

JoVE Journal

Chemistry

11.0K 次观看.  University of Groningen. 电寻址，高纵横比（> 1000:1）制造金属纳米线的差距可以使用铝和银的牺牲层自组装单分子膜为模板的单纳米分离。这些纳隙结构没有一个干净的房间或任何边缘光刻称为nanoskiving的的一种形式由光或电子束光刻工艺制作。

视频: Fabricating Nanogaps by Nanoskiving

Anticancer Metal Complexes: Synthesis and Cytotoxicity Evaluation by the MTT Assay

Titanium (IV) and vanadium (V) complexes are highly potent anticancer agents. A challenge in their synthesis refers to their hydrolytic instability; therefore their preparation should be conducted under an inert atmosphere. Evaluation of the anticancer activity of these complexes can be achieved by the MTT assay.
The MTT assay is a colorimetric viability assay based on enzymatic reduction of the MTT molecule to formazan when it is exposed to viable cells. The outcome of the reduction is a color change of the MTT molecule. Absorbance measurements relative to a control determine the percentage of remaining viable cancer cells following their treatment with varying concentrations of a tested compound, which is translated to the compound anticancer activity and its IC50 values. The MTT assay is widely common in cytotoxicity studies due to its accuracy, rapidity, and relative simplicity.
Herein we present a detailed protocol for the synthesis of air sensitive metal based drugs and cell viability measurements, including preparation of the cell plates, incubation of the compounds with the cells, viability measurements using the MTT assay, and determination of IC50 values.

A method for synthesis of air-sensitive titanium and vanadium anticancer agents is described, along with the evaluation of their cytotoxic activity towards human cancer cell line by the MTT Assay.

抗癌金属配合物的合成与细胞毒性评价MTT法

Titanium (IV) and vanadium (V) complexes are highly potent anticancer agents. A challenge in their synthesis refers to their hydrolytic instability; ...

科研

化学

Preparing Adherent Cells for X-ray Fluorescence Imaging by Chemical Fixation

X-ray fluorescence imaging allows us to non-destructively measure the spatial distribution and concentration of multiple elements simultaneously over large or small sample areas. It has been applied in many areas of science, including materials science, geoscience, studying works of cultural heritage, and in chemical biology. In the case of chemical biology, for example, visualizing the metal distributions within cells allows us to study both naturally-occurring metal ions in the cells, as well as exogenously-introduced metals such as drugs and nanoparticles. Due to the fully hydrated nature of nearly all biological samples, cryo-fixation followed by imaging under cryogenic temperature represents the ideal imaging modality currently available. However, under the circumstances that such a combination is not easily accessible or practical, aldehyde based chemical fixation remains useful and sometimes inevitable. This article describes in as much detail as possible in the preparation of adherent mammalian cells by chemical fixation for X-ray fluorescent imaging.

Here, we present a protocol on how to determine the quantity and distribution of metals in a sample using synchrotron X-ray fluorescence. We focus on adherent cells, and describe the chemical fixation method to prepare this sample. We then describe how to mount and image the sample using synchrotron X-rays.

化学固定准备贴壁细胞的X射线荧光成像

X-ray fluorescence imaging allows us to non-destructively measure the spatial distribution and concentration of multiple elements simultaneously over ...

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

Enzyme catalysis evolved in an aqueous environment. The influence of solvent dynamics on catalysis is, however, currently poorly understood and usually neglected. The study of water dynamics in enzymes and the associated thermodynamical consequences is highly complex and has involved computer simulations, nuclear magnetic resonance (NMR) experiments, and calorimetry. Water tunnels that connect the active site with the surrounding solvent are key to solvent displacement and dynamics. The protocol herein allows for the engineering of these motifs for water transport, which affects specificity, activity and thermodynamics. By providing a biophysical framework founded on theory and experiments, the method presented herein can be used by researchers without previous expertise in computer modeling or biophysical chemistry. The method will advance our understanding of enzyme catalysis on the molecular level by measuring the enthalpic and entropic changes associated with catalysis by enzyme variants with obstructed water tunnels. The protocol can be used for the study of membrane-bound enzymes and other complex systems. This will enhance our understanding of the importance of solvent reorganization in catalysis as well as provide new catalytic strategies in protein design and engineering.

Channels for the transportation of water molecules in enzymes influence active site solvation and catalysis. Herein we present a protocol for the engineering of these additional catalytic motifs based on in silico computer modeling and experiments. This will enhance our understanding of the influence of solvent dynamics on enzyme catalysis.

揭开熵率的加速溶剂动力学的膜酶诱导

Enzyme catalysis evolved in an aqueous environment. The influence of solvent dynamics on catalysis is, however, currently poorly understood and usually ...

Sulfate Separation by Selective Crystallization with a Bis-iminoguanidinium Ligand

A simple and effective method for selective sulfate separation from aqueous solutions by crystallization with a bis-guanidinium ligand, 1,4-benzene-bis(iminoguanidinium) (BBIG), is demonstrated. The ligand is synthesized as the chloride salt (BBIG-Cl) by in situ imine condensation of terephthalaldehyde with aminoguanidinium chloride in water, followed by crystallization as the sulfate salt (BBIG-SO4). Alternatively, BBIG-Cl is synthesized ex situ in larger scale from ethanol. The sulfate separation ability of the BBIG ligand is demonstrated by selective and quantitative crystallization of sulfate from seawater. The ligand can be recycled by neutralization of BBIG-SO4 with aqueous NaOH and crystallization of the neutral bis-iminoguanidine, which can be converted back into BBIG-Cl with aqueous HCl and reused in another separation cycle. Finally, 35S-labeled sulfate and &#946; liquid scintillation counting are employed for monitoring the sulfate concentration in solution. Overall, this protocol will instruct the user in the necessary skills to synthesize a ligand, employ it in the selective crystallization of sulfate from aqueous solutions, and quantify the separation efficiency.

A protocol for in situ aqueous synthesis of a bis(iminoguanidinium) ligand and its utilization in selective separation of sulfate is presented.

分离硫酸选择性结晶用双 -  iminoguanidinium配体

A simple and effective method for selective sulfate separation from aqueous solutions by crystallization with a bis-guanidinium ligand, ...

DNA-magnetic Particle Binding Analysis by Dynamic and Electrophoretic Light Scattering

Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the evaluation of DNA-magnetic particles binding via dynamic light scattering (DLS) and electrophoretic light scattering (ELS). Analysis by DLS provides valuable information on the physicochemical properties of particles including particle size, polydispersity, and zeta potential. The latter describes the surface charge of the particle which plays major role in electrostatic binding of materials such as DNA. Here, a comparative analysis exploits three chemical modifications of nanoparticles and microparticles and their effects on DNA binding and elution. Chemical modifications by branched polyethylenimine, tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane are investigated. Since DNA exhibits a negative charge, it is expected that zeta potential of particle surface will decrease upon binding of DNA. Forming of clusters should also affect particle size. In order to investigate the efficiency of these particles in isolation and elution of DNA, the particles are mixed with DNA in low pH (~6), high ionic strength and dehydration environment. Particles are washed on magnet and then DNA is eluted by Tris-HCl buffer (pH = 8). DNA copy number is estimated using quantitative polymerase chain reaction (PCR). Zeta potential, particle size, polydispersity and quantitative PCR data are evaluated and compared. DLS is an insightful and supporting method of analysis that adds a new perspective to the process of screening of particles for DNA isolation.

This protocol describes the synthesis of magnetic particles and evaluation of their DNA-binding properties via dynamic and electrophoretic light scattering. This method focuses on monitoring changes in particle size, their polydispersity and zeta potential of particle surface which play major role in binding of materials such as DNA.

动态和电泳光散射的 DNA-磁性颗粒结合分析

Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity and convenient colorimetric readout. However, high densities of nanoparticles are typically needed for detection. The available synthesis-based enhancement protocols are either limited to gold and silver nanoparticles or rely on precise enzymatic control and optimization. Here, we present a protocol to enhance the colorimetric readout of gold, silver, silica, and iron oxide nanoprobes. It was observed that the colorimetric signal can be improved by up to a 10000-fold factor. The basis for such signal enhancement strategies is the chemical reduction of Au3+ to Au0. There are several chemical reactions that enable the reduction of Au3+ to Au0. In the protocol, Good&#39;s buffers and H2O2 are used and it is possible to favor the deposition of Au0 onto the surface of existing nanoprobes, in detriment of the formation of new gold nanoparticles. The protocol consists of the incubation of the microarray with a solution consisting of chloroauric acid and H2O2 in 2-(N-morpholino)ethanesulfonic acid pH 6 buffer following the nanoprobe-based detection assay. The enhancement solution can be applied to paper and glass-based sensors. Moreover, it can be used in commercially available immunoassays as demonstrated by the application of the method to a commercial allergen microarray. The signal development requires less than 5 min of incubation with the enhancement solution and the readout can be assessed by naked eye or low-end image acquisition devices such as a table-top scanner or a digital camera.

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles.

原位金纳米粒子合成快速 Nanoprobe 信号增强

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...

Fabricating High-viscosity Droplets using Microfluidic Capillary Device with Phase-inversion Co-flow Structure

The generation of monodisperse droplets with high viscosity has always been a challenge in droplet microfluidics. Here, we demonstrate a phase-inversion co-flow device to generate uniform high-viscosity droplets in a low-viscosity fluid. The microfluidic capillary device has a common co-flow structure with its exit connecting to a wider tube. Elongated droplets of the low-viscosity fluid are first encapsulated by the high-viscosity fluid in the co-flow structure. As the elongated low-viscosity droplets flow through the exit, which is treated to be wetted by the low-viscosity fluid, phase inversion is then induced by the adhesion of the low viscosity droplets to the tip of the exit, which results in the subsequent inverse encapsulation of the high-viscosity fluid. The size of the resultant high-viscosity droplets can be adjusted by changing the flow rate ratio of the low-viscosity fluid to the high-viscosity fluid. We demonstrate several typical examples of the generation of high-viscosity droplets with a viscosity up to 11.9 Pas, such as glycerol, honey, starch, and polymer solution. The method provides a simple and straightforward approach to generate monodisperse high-viscosity droplets, which may be used in a variety of droplet-based applications, such as materials synthesis, drug delivery, cell assay, bioengineering, and food engineering.

A phase-inversion co-flow device is demonstrated to generate monodisperse high-viscosity droplets above 1 Pas, which is difficult to realize in droplet microfluidics.

反相共流微流控毛细管装置制备高粘度水滴的方法

The generation of monodisperse droplets with high viscosity has always been a challenge in droplet microfluidics. Here, we demonstrate a phase-inversion ...

A Rhodopsin Transport Assay by High-Content Imaging Analysis

Rhodopsin misfolding mutations lead to rod photoreceptor death that is manifested as autosomal dominant retinitis pigmentosa (RP), a progressive blinding disease that lacks effective treatment. We hypothesize that the cytotoxicity of the misfolded rhodopsin mutant can be alleviated by pharmacologically stabilizing the mutant rhodopsin protein. The P23H mutation, among the other Class II rhodopsin mutations, encodes a structurally unstable rhodopsin mutant protein that is accumulated in the endoplasmic reticulum (ER), whereas the wild type rhodopsin is transported to the plasma membrane in mammalian cells. We previously performed a luminescence-based high-throughput screen (HTS) and identified a group of pharmacological chaperones that rescued the transport of the P23H rhodopsin from ER to the plasma membrane. Here, using an immunostaining method followed by a high-content imaging analysis, we quantified the mutant rhodopsin protein amount in the whole cell and on the plasma membrane. This method is informative and effective to identify true hits from false positives following HTS. Additionally, the high-content image analysis enabled us to quantify multiple parameters from a single experiment to evaluate the pharmacological properties of each compound. Using this assay, we analyzed the effect of 11 different compounds towards six RP associated rhodopsin mutants, obtaining a 2-D pharmacological profile for a quantitative and qualitative understanding about the structural stability of these rhodopsin mutants and efficacy of different compounds towards these mutants.

Here, we described a high-content imaging method to quantify the transport of rhodopsin mutants associated with retinitis pigmentosa. A multiple-wavelength scoring analysis was used to quantify rhodopsin protein on the cell surface or in the whole cell.

高含量成像分析研究。

Rhodopsin misfolding mutations lead to rod photoreceptor death that is manifested as autosomal dominant retinitis pigmentosa (RP), a progressive blinding ...

Photodeposition of Pd onto Colloidal Au Nanorods by Surface Plasmon Excitation

A protocol is described to photocatalytically guide Pd deposition onto Au nanorods (AuNR) using surface plasmon resonance (SPR). Excited plasmonic hot electrons upon SPR irradiation drive reductive deposition of Pd on colloidal AuNR in the presence of [PdCl4]2-. Plasmon-driven reduction of secondary metals potentiates covalent, sub-wavelength deposition at targeted locations coinciding with electric field &#8220;hot-spots&#8221; of the plasmonic substrate using an external field (e.g., laser). The process described herein details a solution-phase deposition of a catalytically-active noble metal (Pd) from a transition metal halide salt (H2PdCl4) onto aqueously-suspended, anisotropic plasmonic structures (AuNR). The solution-phase process is amenable to making other bimetallic architectures. Transmission UV-vis monitoring of the photochemical reaction, coupled with ex situ XPS and statistical TEM analysis, provide immediate experimental feedback to evaluate properties of the bimetallic structures as they evolve during the photocatalytic reaction. Resonant plasmon irradiation of AuNR in the presence of [PdCl4]2- creates a thin, covalently-bound Pd0 shell without any significant dampening effect on its plasmonic behavior in this representative experiment/batch. Overall, plasmonic photodeposition offers an alternative route for high-volume, economical synthesis of optoelectronic materials with sub-5 nm features (e.g., heterometallic photocatalysts or optoelectronic interconnects).

A protocol for anisotropic photodeposition of Pd onto aqueously-suspended Au nanorods via localized surface plasmon excitation is presented.

通过表面等离子体激发将 Pd 光沉积到胶体 Au 纳米棒上

A protocol is described to photocatalytically guide Pd deposition onto Au nanorods (AuNR) using surface plasmon resonance (SPR). Excited plasmonic hot ...

Obtention of Giant Unilamellar Hybrid Vesicles by Electroformation and Measurement of their Mechanical Properties by Micropipette Aspiration

Giant vesicles obtained from phospholipids and copolymers can be exploited in different applications: controlled and targeted drug delivery, biomolecular recognition within biosensors for diagnosis, functional membranes for artificial cells, and development of bioinspired micro/nano-reactors. In all of these applications, the characterization of their membrane properties is of fundamental importance. Among existing characterization techniques, micropipette aspiration, pioneered by E. Evans, allows the measurement of mechanical properties of the membrane such as area compressibility modulus, bending modulus and lysis stress and strain. Here, we present all the methodologies and detailed procedures to obtain giant vesicles from the thin film of a lipid or copolymer (or both), the manufacturing and surface treatment of micropipettes, and the aspiration procedure leading to the measurement of all the parameters previously mentioned.

The goal of the protocol is to reliably measure membrane mechanical properties of giant vesicles by micropipette aspiration.

通过电形成和微移液器测量其机械性能，对巨型Unilamellar混合囊泡的吸收

Giant vesicles obtained from phospholipids and copolymers can be exploited in different applications: controlled and targeted drug delivery, biomolecular ...