这项工作是由美国国立卫生研究院资助R01 EB008101和R01 CA103830部分支持。

1，仪器及玻璃制品的制备<ol><li>穿戴适当的防护设备， 即一个白大褂，一次性手套和护目镜。 </li><li>圆底烧瓶连接到冷凝器和浸在硅油浴与温度计的温度监测。放的热量下的油浴中（ 图1）的来源（ 例如，热板）。使用能够测量温度高于260℃的温度计。 </li></ol> 2，初级合成的混合磁等离激元纳米粒子<ol><li>使磁芯纳米粒子<ol><li>加入353.2毫克（1毫摩尔），铁（III）乙酰丙酮化物，将1ml（2毫摩尔）油酸，将1ml（2毫摩尔）的油胺，1.292克（5毫摩尔）1,2 - 十六烷和10ml苯醚为圆汉字瓶。 </li><li>大力使用磁力搅拌棒，加热至250-260℃，回流1小时搅拌混合。然后，等待溶液冷却至室温。确保温度低于260℃，以防止苯基醚的沸点和防止突发的反应混合物从圆底烧瓶中，向冷凝器。 注意：反应混合物是非常热和化学物质可能引起刺激。操作必须在通风橱和穿戴适当的个人防护装备。保证充分的通风的油浴。 注：合成的磁性纳米粒子的过程中，油浴中保持在250-260ºC温度1小时。在原则上，派热克斯玻璃皿可以被用于此目的。然而，最大的连续温度为硼硅玻璃是〜260ºC根据供应商的信息。因此，金属容器提供的，因为它可以承受更高的温度，并在多个运行持续时间越长，反应更安全的选择。 </li></ol></li><li>沉积金壳上的磁性纳米粒子核<ol><li>添加411.5毫克（1.1毫摩尔）金王牌的tate，加入0.25ml（0.75毫摩尔）油酸，将1.5ml（3.0毫摩尔）的油胺，775.3毫克（3毫摩尔）1,2 - 十六烷和15ml苯醚至圆底烧瓶中。 </li><li>加入5 mL悬浮磁性纳米颗粒的步骤2.1。将反应混合物加热至180℃并保持在回流下搅拌1小时。等待该溶液冷却至室温。 </li><li>加入50毫升乙醇中，以沉淀混合初级纳米颗粒，然后离心以3250×g的15分钟。 </li><li>用水浴超声悬浮于25ml正己烷沉淀。加25毫升乙醇中，以沉淀出主混合纳米粒子。离心机在3250×g离心15分钟，重悬沉淀在己烷中。重复此步骤三次。 </li><li>干燥在真空干燥器中O 2 / N的沉淀主要混合纳米粒子。确认该颗粒是完全干燥的。 </li></ol></li></ol> 3，混合动力磁等离子团簇合成和大小分离<ol><li&gt;重悬于5毫克干初级混合纳米粒子在1毫升己烷用水浴超声。超声处理的纳米颗粒悬浮液，直到没有可见沉淀物的存在。 </li><li>从步骤加入该溶液3.1至10ml的十二烷基硫酸钠（2.8毫克/毫升）水溶液中以附帽20毫升玻璃小瓶中。添加的主混合纳米颗粒滴悬浮液一滴地避免了两个阶段的下一个步骤之前混合。 </li><li>超声处理在水浴超声处理2小时的两相溶液中，然后通过加热的水浴中，在80℃下进行10分钟。等待该溶液冷却至室温。 <ol><li>填充水的超声浴中的操作电平线。居中在超声处理浴中的玻璃小瓶中。一种乳液形式立即在两相之间。摇通过超声处理的开始之后手的两相溶液;这有利于含伯混合纳米颗粒的相和底部的水相之间的混合。 注：请注意THAT中的超声仪2小时手术后会热起来。 </li></ol></li><li>离心反应混合纳米簇悬浮液在100×g离心30分钟。同时收集沉淀物和上清液。悬浮于0.1 mM柠檬酸钠沉淀物在10分钟超声。纳米团簇的预期大小是直径〜180纳米。 </li><li>步骤3.3将上清转移到一个新的锥形管。 </li><li>离心从步骤3.4的悬浮液在400×g离心30分钟。同时收集沉淀物和上清液。悬浮于0.1 mM柠檬酸钠沉淀物在10分钟超声。纳米团簇的预期大小是直径〜130纳米。 </li><li>步骤3.5将上清转移到一个新的锥形管。 </li><li>离心从步骤3.6的悬浮液在1500×g离心30分钟。收集0.1 mM柠檬酸钠沉淀重悬在10分钟超声。纳米团簇的预期大小是直径约90纳米。 </li><li>加入300 _6;升纳米簇悬浮液至96孔微板读数器，用于测量紫外 - 可见 - 近红外吸收光谱。下降10微升悬浮液中纳米团簇上的碳包覆铜网格TEM成像。 </li></ol> 4，共轭单克隆抗体的制备纳米团簇<ol><li>制备100μl的单克隆抗体溶液（1毫克/毫升）的PBS，pH为7.2， 例如 ，抗表皮生长因子受体2（HER2）的抗体或抗表皮生长因子受体1（EGFR）抗体。 </li><li>步骤4.1抗体溶液中加入3.9毫升4毫米肝素钠，pH值7.2。离心机通过10 K截留分子量离心过滤的解决方案，在3,250×g离心20分钟，在8℃。重悬的抗体在4毫米的HEPES，pH值7.2，于100微升的最终体积。 注意：此步骤是，以取代在与HEPES的抗体溶液的原始媒体。 </li><li>加入10微升100mM的NaIO 4至100μl的抗体溶液。覆盖所述反应小瓶用一个人的使用轨道摇床uminum箔在RT，搅拌30分钟。 </li><li>淬灭，加入500μl的1×PBS中的反应。 </li><li>加入2微升46.5毫米的连接器解决方案（dithiolaromatic PEG6-CONHNH 2）抗体溶液从步骤4.4振摇1小时在室温。 </li><li>过滤器采用10 K截留分子量离心过滤器的解决方案，在3,250×g离心20分钟，在8℃。重新悬浮于1×PBS中的抗体至100μl导致的〜1毫克/毫升抗体浓度的最终体积。 </li><li>从步骤4.6（1毫克/毫升）120分钟在室温下混合100微升纳米簇悬浮在OD〜1.0用1μl的修饰抗体。 </li><li>加入10微升10 -3 M 5 kDa的巯基聚乙二醇，振摇15分钟在室温。 </li><li>离心反应溶液在830×g下3分钟。弃上清，重悬在100微升2％的泥沙w / v的5 kDa的PEG在PBS，pH值为7.2。 </li><li>测定抗体偶联的纳米团簇的吸收光谱，并比较吨他吸光度裸露的纳米团簇的光谱。期待一对夫妇的结合后，纳米红移。 </li><li>如果纳米粒子团聚物由显著移与在红色-近红外区域的增加OD所示，增加硫醇的PEG的浓度为5×10 -3 M。此外，增加了孵育时间与巯基的PEG至30分钟，并降低了离心速度在200 XG增量。 </li><li>对于癌症细胞标记测试中，从步骤4.9到癌症细胞悬浮液在中等或1×PBS中（1毫升〜10 6个细胞）中添加的抗体缀合的纳米颗粒，并混合60分钟。 </li></ol>

<ol>
	<li>Bigall, N. C., Parak, W. J., Dorfs, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent,+magnetic+and+plasmonic&#8212;Hybrid+multifunctional+colloidal+nano+objects.">Fluorescent, magnetic and plasmonic&#8212;Hybrid multifunctional colloidal nano objects.</a> Nano Today. 7, 282-296 (2012).</li><li>Gautier, J., Allard-Vannier, E., Herve-Aubert, K., Souce, M., Chourpa, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+strategies+of+hybrid+metallic+nanoparticles+for+theragnostic+applications.">Design strategies of hybrid metallic nanoparticles for theragnostic applications.</a> Nanotechnology. 24, 432002 (2013).</li><li>Wei, Q., Wei, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+imaging+with+dynamic+contrast+agents.">Optical imaging with dynamic contrast agents.</a> Chemistry. 17, 1080-1091 (2011).</li><li>Aaron, J. 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=Increased+optical+contrast+in+imaging+of+epidermal+growth+factor+receptor+using+magnetically+actuated+hybrid+gold/iron+oxide+nanoparticles.">Increased optical contrast in imaging of epidermal growth factor receptor using magnetically actuated hybrid gold/iron oxide nanoparticles.</a> Optics express. 14, 12930-12943 (2006).</li><li>Song, H. -. M., Wei, Q., Ong, Q. K., Wei, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasmon-resonant+nanoparticles+and+nanostars+with+magnetic+cores:+synthesis+and+magnetomotive+imaging.">Plasmon-resonant nanoparticles and nanostars with magnetic cores: synthesis and magnetomotive imaging.</a> ACS nano. 4, 5163-5173 (2010).</li><li>Qu, 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=Magneto-photo-acoustic+imaging.">Magneto-photo-acoustic imaging.</a> Biomedical optics express. 2, 385-396 (2011).</li><li>Jin, Y., Jia, C., Huang, S. -. W., Donnell O&amp;39, M., Gao, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multifunctional+nanoparticles+as+coupled+contrast+agents.">Multifunctional nanoparticles as coupled contrast agents.</a> Nature communications. 1, 41 (2010).</li><li>Wu, C. -. 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=Versatile+Immunomagnetic+Nanocarrier+Platform+for+Capturing+Cancer+Cells.">Versatile Immunomagnetic Nanocarrier Platform for Capturing Cancer Cells.</a> ACS. 7, 8816-8823 (2013).</li><li>Galanzha, E. I., 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=In+vivo+magnetic+enrichment+and+multiplex+photoacoustic+detection+of+circulating+tumour+cells.">In vivo magnetic enrichment and multiplex photoacoustic detection of circulating tumour cells.</a> Nature nanotechnology. 4, 855-860 (2009).</li><li>Larson, T. A., Bankson, J., Aaron, J., Sokolov, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hybrid+plasmonic+magnetic+nanoparticles+as+molecular+specific+agents+for+MRI/optical+imaging+and+photothermal+therapy+of+cancer+cells.">Hybrid plasmonic magnetic nanoparticles as molecular specific agents for MRI/optical imaging and photothermal therapy of cancer cells.</a> Nanotechnology. 18, 325101 (2007).</li><li>Yu, 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=Dumbbell-like+bifunctional+Au-Fe3O4+nanoparticles.">Dumbbell-like bifunctional Au-Fe3O4 nanoparticles.</a> Nano letters. 5, 379-382 (2005).</li><li>Wang, L., 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=Monodispersed+core-shell+Fe3O4@Au+nanoparticles.">Monodispersed core-shell Fe3O4@Au nanoparticles.</a> The journal of physical chemistry. B. 109, 21593-21601 (2005).</li><li>Wang, H., Brandl, D. W., Le, F., Nordlander, P., Halas, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanorice:+a+hybrid+plasmonic+nanostructure.">Nanorice: a hybrid plasmonic nanostructure.</a> Nano letters. 6, 827-832 (2006).</li><li>Hu, X., 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=Trapping+and+Photoacoustic+Detection+of+CTCs+at+the+Single+Cell+per+Milliliter+Level+with+Magneto&#8208;Optical+Coupled+Nanoparticles.">Trapping and Photoacoustic Detection of CTCs at the Single Cell per Milliliter Level with Magneto&#8208;Optical Coupled Nanoparticles.</a> Small. 9, 2046-2052 (2013).</li><li>Truby, R. L., Emelianov, S. Y., Homan, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ligand-mediated+self-assembly+of+hybrid+plasmonic+and+superparamagnetic+nanostructures.">Ligand-mediated self-assembly of hybrid plasmonic and superparamagnetic nanostructures.</a> Langmuir. 29, 2465-2470 (2013).</li><li>Bai, 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=A+Versatile+Bottom&#8208;up+Assembly+Approach+to+Colloidal+Spheres+from+Nanocrystals.">A Versatile Bottom&#8208;up Assembly Approach to Colloidal Spheres from Nanocrystals.</a> Angewandte Chemie International Edition. 46, 6650-6653 (2007).</li></ol>

作者宣称，他们没有竞争的经济利益。

在成功地合成了磁 - 等离子体激元纳米簇的关键步骤包括：拍摄高度单分散的初级金壳/氧化铁核心纳米颗粒和引导的一次粒子的自组装成纳米团簇。初级粒子和表面活性剂之间的摩尔比在确定的纳米团簇的尺寸分布中起重要作用。初级纳米颗粒的非均匀的粒度分布的磁 - 等离子体激元纳米簇组装期间导致形成大的聚集体。此外，纳米簇形成的微乳状液的方法依赖于两亲性表面活性剂：疏水性尾部...

混合纳米粒子由不同的材料具有不同的理化性质，可在医学领域的应用，包括多模式分子影像学，治疗和交付的监控，新的筛选和诊断分析1-3打开新的机会。等离激元和磁特性的单个纳米颗粒的组合是特别感兴趣的，因为它提供了与等离子体激元共振和响应相关联的磁场很强的光散射和吸收截面。例如，磁-等离子体激元纳米颗粒，使用了经由外部电磁铁3-5施加时间信号调制，以增加在标记细胞的暗场成像的对比度。磁光声成像，在磁等离激元纳米粒子能在对比度和信后台大鼠很大的改进 - 最近，类似的原则在新的成像方法的发展施加IO 6,7。它还表明，该混合纳米粒子可用于同时捕获和检测全血中和体内 8,9循环肿瘤细胞。此外，磁-等离子体激元纳米颗粒是有希望的治疗诊断剂，其可用于分子特定的光学和磁共振成像结合癌细胞10的光热治疗。 有几种方法进行了探讨合成磁等离激元纳米粒子。例如，Yu 等人利用分解和Fe（CO）的氧化5金纳米粒子，形成哑铃状的双功能金， 四氧化三铁纳米粒子11。王等人已经用热分解方法12合成的涂覆金的氧化铁纳米颗粒。一些其他方法依赖于涂层聚合物或胺官能分子上磁心纳米颗粒随后AG的沉积旧壳到聚合物表面以产生杂种粒子7,13。此外，氧化铁纳米颗粒通过静电相互作用或发生化学反应14,15连接到金纳米棒。虽然这些方法中，得到磁 - 等离子体激元纳米结构，它们兼顾到磁电浆组合的某种程度的属性，例如光吸收在近红外（NIR）窗口或一个强大的磁力矩这两者都是在生物医学应用中是非常理想的。例如，哑铃金- 四氧化三铁纳米粒子在520nm处，限制其在体内的效用，由于高浊度的组织在这个光谱范围内的等离子体共振峰。此外，通过当前的协议所产生的磁-等离子体激元纳米颗粒被局限于一个11或少数（小于10），14,15超顺磁性基团（ 例如 ，氧化铁纳米颗粒）是显著小于可能是ACHieved在密密麻麻的纳米结构。例如，密密麻麻的直径60纳米的球形纳米颗粒可以包含上千6纳米超顺磁性纳米颗粒的数量级上。因此，存在对于改善混合纳米粒子的磁特性有很大空间。此外，一些先前所描述的协议是相对复杂的，需要以避免合成14,15中粒子的凝集仔细优化。 在这里，我们描述了一个协议，用于合成磁等离激元纳米粒子具有较强的磁矩和强大的近红外吸收，解决了目前先进的主要限制。合成有其油包水微乳液法16渊源。它是基于来自一个更小的一次粒子的纳米粒子所需的尺寸的组件。这种方法已被成功地用于生产由单一材料的纳米结构，例如金，铁氧化物和半导体的pri玛丽颗粒16。我们扩展它来合成磁电浆纳米颗粒的通过，第一，使得6纳米直径的金壳/氧化铁芯颗粒，然后，装配主杂种颗粒进入最后的球形纳米结构。组装一次颗粒的纳米团簇，不仅使增强组分的纳米颗粒，如实现更强的磁力矩，同时保持超顺磁性的特性，但还需要单独的纳米颗粒从而形成从构成的纳米粒子不存在的新的特性，如强之间的相互作用的优点光吸收的近红外窗口。这个协议产生混合纳米粒子与磁场和等离激元的功能的高密度。后一次粒子合成出，我们的方法本质上是一个简单的单罐反应。整体的等离子体共振的强度和磁矩是由一些初级粒子和，疗法的确定安伏，可以根据用途容易地优化。此外，我们还制定了抗体偶联过程的混合纳米粒子的各种生物医学应用程序需要特定的分子靶向。抗体通过与Fc部分离开Fab部分，它负责抗原结合可用于靶向附着。

<table border="0" cellpadding="0" cellspacing="0" width="738">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Name of Material/ Equipment</td>
			<td style="width:101px;">Company</td>
			<td style="width:139px;">Catalog Number</td>
			<td style="width:247px;">Comments/Description</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:253px;">PYREX 50mL Round Bottom Boiling Flask with Short Neck &#38; 24/40 [ST] Joint</td>
			<td>Corning</td>
			<td style="width:139px;">4320A-50</td>
			<td>Thermal decomposition reaction</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:253px;">PYREX 41 x 300mm 5-Bulb Allihn Condenser with 24/40 [ST] Outer/Inner Joints</td>
			<td>Corning</td>
			<td style="width:139px;">2480-300</td>
			<td>Thermal decomposition reaction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Silicone Oil</td>
			<td>Fisher</td>
			<td style="width:139px;">S159-500</td>
			<td>Oil bath</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Hot Plate Stirrer</td>
			<td>Corning</td>
			<td style="width:139px;">PC-351</td>
			<td>Heat the reacton with stirring function</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Thermometer</td>
			<td>ThermoWorks</td>
			<td style="width:139px;">221-092</td>
			<td>Measure temperature</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:253px;">Iron (III) Acetylacetonate</td>
			<td>Fisher</td>
			<td style="width:139px;">AC11913-0250</td>
			<td rowspan="7" style="width:247px;">Materials for primary hybrid nanoparticles synthesis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Oleic Acid 99%</td>
			<td>Fisher</td>
			<td style="width:139px;">A195-500</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Gold (III) Acetate</td>
			<td>Fisher</td>
			<td style="width:139px;">AA3974206</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Hexane</td>
			<td>Fisher</td>
			<td style="width:139px;">H292-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Phenyl Ether 99%</td>
			<td>Fisher</td>
			<td style="width:139px;">AC13060-0025</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1,2-Hexadecanediol 90%</td>
			<td>Sigma</td>
			<td style="width:139px;">213748-50G</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Oleylamine 70%</td>
			<td>Sigma</td>
			<td style="width:139px;">O7805-100G</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Sodium Dodecyl Sulfate</td>
			<td style="width:101px;">Fisher</td>
			<td style="width:139px;">BP166-100</td>
			<td rowspan="2" style="width:247px;">Cluster synthesis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Sodium Citrate Dihydrate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:139px;">W302600</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:253px;">Monoclonal Anti-EGF Receptor Antibody</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:139px;">E2156</td>
			<td style="width:247px;">Cell labeling specificity test</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Monoclonal Anti-HER2 Antibody</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:139px;">AMAB90627</td>
			<td style="width:247px;">Cell labeling specificity test</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Sodium Periodate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:139px;">311448</td>
			<td style="width:247px;">Oxidate Fc region of antibodies</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:253px;">Dithiolaromatic PEG6-CONHNH2</td>
			<td style="width:101px;">SensoPath Technologies</td>
			<td style="width:139px;">SPT-0014B</td>
			<td style="width:247px;">Heterofunctional linker for antibody conjugation to nanoclusters</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:253px;">Methoxy-PEG-thiol, 5k</td>
			<td style="width:101px;">Creative PEGworks</td>
			<td style="width:139px;">PLS-604</td>
			<td style="width:247px;">Passivate the remaining gold surface after antibody conjugation</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:253px;">Amicon Ultra-4 Centrifugal Filter Unit with Ultracel-10 membrane</td>
			<td style="width:101px;">Millipore</td>
			<td style="width:139px;">UFC801008</td>
			<td style="width:247px;">Protien purification</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">HEPES</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:139px;">H3375</td>
			<td style="width:247px;">Buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">PBS, 1X Solution</td>
			<td style="width:101px;">Fisher</td>
			<td style="width:139px;">BP2438-20</td>
			<td style="width:247px;">Buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">UV&#8722;vis Spectroscopy</td>
			<td style="width:101px;">BioTek&#160;</td>
			<td style="width:139px;">Synergy HT</td>
			<td style="width:247px;">Obtain spectrum</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Centrifuge</td>
			<td>Eppendorf</td>
			<td>5810R</td>
			<td>Separation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transmission Electron Microscope</td>
			<td>FEI</td>
			<td>TECNAI G2 F20 X-TWIN</td>
			<td>Obtain morphology of nanostructures</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Upright microscope</td>
			<td>Leica</td>
			<td>DM6000</td>
			<td>Obtain dark-field images</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sonicator</td>
			<td>Branson</td>
			<td>1510</td>
			<td>Sonication</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">Carbon Film 300 Mesh Grid</td>
			<td>EMS</td>
			<td style="width:139px;">CF300-Cu</td>
			<td>TEM imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:253px;">96-well Plate</td>
			<td>Corning</td>
			<td style="width:139px;">09-761-145</td>
			<td>UV-vis reading plate</td>
		</tr>
	</tbody>
</table>

synthesis of immunotargeted magneto-plasmonic nanoclusters

磁性和电浆特性结合在一个单一的纳米颗粒提供了协同作用是有利的，在许多生物医学应用，包括在新的磁成像方式的对比度增强，同时捕获和检测（检验中心）的循环肿瘤细胞，以及多峰分子成像结合光热治疗的癌细胞。这些应用刺激了协议的发展为合成磁电浆纳米颗粒在近红外（NIR）区域的光吸收率和较强的磁矩的显著兴趣。这里，我们提出了一种新的协议，用于合成这种杂化的纳米颗粒是基于一种油包水微乳液的方法。本文所描述的协议的独特特征是合成各种尺寸从主块也具有磁电浆特性的磁 - 等离子体激元纳米颗粒。这种方法产生的纳米颗粒具有高登sity其中在整个所述纳米颗粒体积均匀分布的磁场和等离子官能团。该混合的纳米颗粒可以通过与Fc部分离开Fab部分，它负责抗原结合可用于靶向附着的抗体很容易地官能化。

甲方案为合成immunotargeted磁电浆纳米簇的示于图2，首先，磁性的Fe 3 O 4的氧化铁纳米颗粒是通过热分解法合成出。然后，一个薄约 1纳米金壳上沉积通过热分解氧化铁核心颗粒。主约 6纳米杂化的纳米颗粒作为种子，以通过利用一个油包水微乳液的方法制造磁-等离子体激元纳米团簇。纳米团簇被官能化的分子特异性靶向的单克隆抗体。 <p class="jove_content...

Watch this Scientific Journal Video about Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters at JoVE.com

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

在这里，我们描述了一个协议，用于合成磁等离激元纳米粒子具有较强的磁矩和强大的近红外（NIR）吸收。该协议还包括抗体缀合，以通过与Fc部分关于各种生物医学应用的需要分子特异性靶向的纳米颗粒。

Immunotargeted的合成磁等离子团簇

Magnetic and plasmonic properties combined in a single nanoparticle provide a synergy that is advantageous in a number of biomedical applications ...

synthesis-of-immunotargeted-magneto-plasmonic-nanoclusters

Research

JoVE Journal

Chemistry

15.2K 次观看.  University of Texas at Austin. 在这里，我们描述了一个协议，用于合成磁等离激元纳米粒子具有较强的磁矩和强大的近红外（NIR）吸收。该协议还包括抗体缀合，以通过与Fc部分关于各种生物医学应用的需要分子特异性靶向的纳米颗粒。 

视频: Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

Synthesis of Indoxyl-glycosides for Detection of Glycosidase Activities

Indoxyl glycosides proved to be valuable and versatile tools for monitoring glycosidase activities. Indoxyls are released by enzymatic hydrolysis and are rapidly oxidized, for example by atmospheric oxygen, to indigo type dyes. This reaction enables fast and easy screening in vivo without isolation or purification of enzymes, as well as rapid tests on agar plates or in solution (e.g., blue-white screening, micro-wells) and is used in biochemistry, histochemistry, bacteriology and molecular biology. Unfortunately the synthesis of such substrates proved to be difficult, due to various side reactions and the low reactivity of the indoxyl hydroxyl function. Especially for glucose type structures low yields were observed. Our novel approach employs indoxylic acid ester as key intermediates. Indoxylic acid esters with varied substitution patterns were prepared on scalable pathways. Phase transfer glycosylations with those acceptors and peracetylated glycosyl halides can be performed under common conditions in high yields. Ester cleavage and subsequent mild silver mediated glycosylation yields the peracetylated indoxyl glycosides in high yields. Finally deprotection is performed according to Zempl&#233;n.

Indoxyl glycosides are well-established and widely used tools for enzyme screening and enzyme activity monitoring. Especially for glucose type structures previous syntheses proved to be challenging and low yielding. Our novel approach employs indoxylic acid esters as precious intermediates to yield a considerable number of indoxyl glycosides in good yields.

的吲哚酚苷的检测糖苷酶活动的合成

Indoxyl glycosides proved to be valuable and versatile tools for monitoring glycosidase activities. Indoxyls are released by enzymatic hydrolysis and are ...

科研

化学

Synthesis and Characterization of Supramolecular Colloids

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical properties for applications in photonics, drug delivery and coating technology. Here we present a new family of colloidal building blocks, coined supramolecular colloids, whose self-assembly is controlled through surface-functionalization with a benzene-1,3,5-tricarboxamide (BTA) derived supramolecular moiety. Such BTAs interact via directional, strong, yet reversible hydrogen-bonds with other identical BTAs. Herein, a protocol is presented that describes how to couple these BTAs to colloids and how to quantify the number of coupling sites, which determines the multivalency of the supramolecular colloids. Light scattering measurements show that the refractive index of the colloids is almost matched with that of the solvent, which strongly reduces the van der Waals forces between the colloids. Before photo-activation, the colloids remain well dispersed, as the BTAs are equipped with a photo-labile group that blocks the formation of hydrogen-bonds. Controlled deprotection with UV-light activates the short-range hydrogen-bonds between the BTAs, which triggers the colloidal self-assembly. The evolution from the dispersed state to the clustered state is monitored by confocal microscopy. These results are further quantified by image analysis with simple routines using ImageJ and Matlab. This merger of supramolecular chemistry and colloidal science offers a direct route towards light- and thermo-responsive colloidal assembly encoded in the surface-grafted monolayer.

A protocol for the synthesis and characterization of colloids coated with supramolecular moieties is described. These supramolecular colloids undergo self-assembly upon the activation of the hydrogen-bonds between the surface-anchored molecules by UV-light.

超分子胶体的合成与表征

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Hydroquinone Based Synthesis of Gold Nanorods

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the synthetic methods for the preparation of gold nanorods are still not fully optimized. In this paper we describe a new, highly efficient, two-step protocol based on the use of hydroquinone as a mild reducing agent. Our approach allows the preparation of nanorods with a good control of size and aspect ratio (AR) simply by varying the amount of hexadecyl trimethylammonium bromide (CTAB) and silver ions (Ag+) present in the "growth solution". By using this method, it is possible to markedly reduce the amount of CTAB, an expensive and cytotoxic reagent, necessary to obtain the elongated shape. Gold nanorods with an aspect ratio of about 3 can be obtained in the presence of just 50 mM of CTAB (versus 100 mM used in the standard protocol based on the use of ascorbic acid), while shorter gold nanorods are obtained using a concentration as low as 10 mM.

This paper describes a protocol for the synthesis of gold nanorods, based on the use of hydroquinone as reducing agent, plus the different mechanisms for controlling their size and aspect ratio.

金纳米棒基于对苯二酚合成

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the ...

Synthesis of Hierarchical ZnO/CdSSe Heterostructure Nanotrees

A two-step chemical vapor deposition procedure is here employed to prepare tree-like hierarchical ZnO/CdSSe hetero-nanostructures. The structures are composed of CdSSe branches grown on ZnO nanowires that are vertically aligned on a transparent sapphire substrate. The morphology was measured via scanning electron microscopy. The crystal structure was determined by X-ray powder diffraction analysis. Both the ZnO stem and CdSSe branches have a predominantly wurtzite crystal structure. The mole ratio of S and Se in the CdSSe branches was measured by energy dispersive X-ray spectroscopy. The CdSSe branches result in strong visible light absorption. Photoluminescence (PL) spectroscopy showed that the stem and branches form a type-II heterojunction. PL lifetime measurements showed a decrease in the lifetime of emission from the trees when compared to emission from individual ZnO stems or CdSSe branches and indicate fast charge transfer between CdSSe and ZnO. The vertically aligned ZnO stems provide a direct electron transport pathway to the substrate and allow for efficient charge separation after photoexcitation by visible light. The combination of the abovementioned properties makes ZnO/CdSSe nanotrees promising candidates for applications in solar cells, photocatalysis, and opto-electronic devices.

Here, we prepare and characterize novel tree-like hierarchical ZnO/CdSSe nanostructures, where CdSSe branches are grown on vertically aligned ZnO nanowires. The resulting nanotrees are a potential material for solar energy conversion and other opto-electronic devices.

分层的ZnO / CdSSe异质Nanotrees的合成

A two-step chemical vapor deposition procedure is here employed to prepare tree-like hierarchical ZnO/CdSSe hetero-nanostructures. The structures are ...

Synthesis and Mass Spectrometry Analysis of Oligo-peptoids

Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have been thought of as a type of molecular information storage. Mass spectrometry analysis has been considered the method of choice for sequencing peptoids. Peptoids can be synthesized via solid phase chemistry using a repeating two-step reaction cycle. Here we present a method to manually synthesize oligo-peptoids and to analyze the sequence of the peptoids using tandem mass spectrometry (MS/MS) techniques. The sample peptoid is a nonamer consisting of alternating N-(2-methyloxyethyl)glycine (Nme) and N-(2-phenylethyl)glycine (Npe), as well as an N-(2-aminoethyl)glycine (Nae) at the N-terminus. The sequence formula of the peptoid is Ac-Nae-(Npe-Nme)4-NH2, where Ac is the acetyl group. The synthesis takes place in a commercially available solid-phase reaction vessel. The rink amide resin is used as the solid support to yield the peptoid with an amide group at the C-terminus. The resulting peptoid product is subjected to sequence analysis using a triple-quadrupole mass spectrometer coupled to an electrospray ionization source. The MS/MS measurement produces a spectrum of fragment ions resulting from the dissociation of charged peptoid. The fragment ions are sorted out based on the values of their mass-to-charge ratio (m/z). The m/z values of the fragment ions are compared against the nominal masses of theoretically predicted fragment ions, according to the scheme of peptoid fragmentation. The analysis generates a fragmentation pattern of the charged peptoid. The fragmentation pattern is correlated to the monomer sequence of the neutral peptoid. In this regard, MS analysis reads out the sequence information of the peptoids.

A protocol is described for the manual synthesis of oligo-peptoids followed by sequence analysis by mass spectrometry.

寡 peptoids 的合成及质谱分析

Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have ...

Synthesis of Substrate-Bound Au Nanowires Via an Active Surface Growth Mechanism

Advancing synthetic capabilities is important for the development of nanoscience and nanotechnology. The synthesis of nanowires has always been a challenge, as it requires asymmetric growth of symmetric crystals. Here, we report a distinctive synthesis of substrate-bound Au nanowires. This template-free synthesis employs thiolated ligands and substrate adsorption to achieve the continuous asymmetric deposition of Au in solution at ambient conditions. The thiolated ligand prevented the Au deposition on the exposed surface of the seeds, so the Au deposition only occurs at the interface between the Au seeds and the substrate. The side of the newly deposited Au nanowires is immediately covered with the thiolated ligand, while the bottom facing the substrate remains ligand-free and active for the next round of Au deposition. We further demonstrate that this Au nanowire growth can be induced on various substrates, and different thiolated ligands can be used to regulate the surface chemistry of the nanowires. The diameter of the nanowires can also be controlled with mixed ligands, in which another &#34;bad&#34; ligand could turn on the lateral growth. With the understanding of the mechanism, Au nanowire-based nanostructures can be designed and synthesized.

We report a solution-based method to synthesize substrate-bound Au nanowires. By tuning the molecular ligands used during the synthesis, the Au nanowires can be grown from various substrates with different surface properties. Au nanowire-based nanostructures can also be synthesized by adjusting the reaction parameters.

通过活性表面生长机制合成基片束缚金纳米线

Advancing synthetic capabilities is important for the development of nanoscience and nanotechnology. The synthesis of nanowires has always been a ...

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of their interactions with cell membranes, lipid bilayers, and viruses. The hydrophilic ligands make these particles colloidally stable in aqueous solutions and the combination with hydrophobic ligands creates an amphiphilic particle that can be loaded with hydrophobic drugs, fuse with the lipid membranes, and resist nonspecific protein adsorption. Many of these properties depend on nanoparticle size and the composition of the ligand shell. It is, therefore, crucial to have a reproducible synthetic method and reliable characterization techniques that allow the determination of nanoparticle properties and the ligand shell composition. Here, a one-phase chemical reduction, followed by a thorough purification to synthesize these nanoparticles with diameters below 5 nm, is presented. The ratio between the two ligands on the surface of the nanoparticle can be tuned through their stoichiometric ratio used during synthesis. We demonstrate how various routine techniques, such as transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and ultraviolet-visible (UV-Vis) spectrometry, are combined to comprehensively characterize the physicochemical parameters of the nanoparticles.

Amphiphilic gold nanoparticles can be used in many biological applications. A protocol to synthesize gold nanoparticles coated by a binary mixture of ligands and a detailed characterization of these particles is presented.

两栖金纳米粒子的合成与表征

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of ...

Gold Nanoparticle Synthesis

Gold nanoparticles (Au nanoparticles) that are ~12 nm in diameter were synthesized by rapidly injecting a solution of 150 mg (0.15 mmol) of tetrachloroauric acid in 3.0 g (3.7 mmol, 3.6 mL) of oleylamine (technical grade) and 3.0 mL of toluene into a boiling solution of 5.1 g (6.4 mmol, 8.7 mL) of oleylamine in 147 mL of toluene. While boiling and mixing the reaction solution for 2 hours, the color of the reaction mixture changed from clear, to light yellow, to light pink, and then slowly to dark red. The heat was then turned off, and the solution was allowed to gradually cool down to room temperature for 1 hour. The gold nanoparticles were then collected and separated from the solution using a centrifuge and washed three times; by vortexing and dispersing the gold nanoparticles in 10 mL portions of toluene, and then precipitating the gold nanoparticles by adding 40 mL portions of methanol and spinning them in a centrifuge. The solution was then decanted to remove any remaining byproducts and unreacted starting materials. Drying the gold nanoparticles in a vacuum environment produced a solid black pellet; which could be stored for long periods of time (up to one year) for later use, and then redissolved in organic solvents such as toluene.

A protocol for synthesizing ~12 nm diameter gold nanoparticles (Au nanoparticles) in an organic solvent is presented. The gold nanoparticles are capped with oleylamine ligands to prevent agglomeration. The gold nanoparticles are soluble in organic solvents such as toluene.

金纳米粒子合成

Gold nanoparticles (Au nanoparticles) that are ~12 nm in diameter were synthesized by rapidly injecting a solution of 150 mg (0.15 mmol) of ...

Construction of Cyclic Cell-Penetrating Peptides for Enhanced Penetration of Biological Barriers

Cancer has been a grand challenge in global health. However, the complex tumor microenvironment generally limits the access of therapeutics to deeper tumor cells, leading to tumor recurrence. To conquer the limited penetration of biological barriers, cell-penetrating peptides (CPPs) have been discovered with excellent membrane translocation ability and have emerged as useful molecular transporters for delivering various cargoes into cells. However, conventional linear CPPs generally show compromised proteolytic stability, which limits their permeability across biological barriers. Thus, the development of novel molecular transporters that can penetrate biological barriers and exhibit enhanced proteolytic stability is highly desired to promote drug delivery efficiency in biomedical applications. We have previously synthesized a panel of short cyclic CPPs with aromatic crosslinks, which exhibited superior permeability in cancer cells and tissues compared to their linear counterparts. Here, a concise protocol is described for the synthesis of the fluorescently labeled cyclic polyarginine R8 peptide and its linear counterpart, as well as key steps for investigating their cell permeability.

This protocol describes the synthesis of cyclic cell-penetrating peptides with aromatic cross-links and the evaluation of their permeability across biological barriers.

构建环状细胞穿透肽以增强生物屏障的穿透力

Cancer has been a grand challenge in global health. However, the complex tumor microenvironment generally limits the access of therapeutics to deeper ...

等离子体金锡合金纳米颗粒的水合成

Aqueous Synthesis of Plasmonic Gold-Tin Alloy Nanoparticles

This protocol describes the synthesis of Au nanoparticle seeds and the subsequent formation of Au-Sn bimetallic nanoparticles. These nanoparticles have potential applications in catalysis, optoelectronics, imaging, and drug delivery. Previously, methods for producing alloy nanoparticles have been time-consuming, require complex reaction conditions, and can have inconsistent results. The outlined protocol first describes the synthesis of approximately 13 nm Au nanoparticle seeds using the Turkevich method. The protocol next describes the reduction of Sn and its incorporation into the Au seeds to generate Au-Sn alloy nanoparticles. The optical and structural characterization of these nanoparticles is described. Optically, prominent localized surface plasmon resonances (LSPRs) are apparent using UV-visible spectroscopy. Structurally, powder X-ray diffraction (XRD) reflects all particles to be less than 20 nm and shows patterns for Au, Sn, and multiple Au-Sn intermetallic phases. Spherical morphology and size distribution are obtained from transmission electron microscopy (TEM) imaging. TEM reveals that after Sn incorporation, the nanoparticles grow to approximately 15 nm in diameter.

作者聚焦：设计可持续的纳米材料以促进合成和元素混合

Here, the synthesis of gold (Au) seeds is described using the Turkevich method. These seeds are then used to synthesize gold-tin alloy (Au-Sn) nanoparticles with tunable plasmonic properties.

This protocol describes the synthesis of Au nanoparticle seeds and the subsequent formation of Au-Sn bimetallic nanoparticles. These nanoparticles have ...