We thank E. Arzt for his support through INM. The research was in part supported by the Leibniz Competition 2014.

<ol>
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The authors have nothing to disclose.

所描述的协议能够在液体中金纳米粒子的茎，包括动态过程的观察。保持器的组装是容易了解的技术。但是，与液体流的TEM保持器工作时，若干方面必须加以考虑。例如，在O形环的硅微芯片或大颗粒的破碎边缘可能导致液体细胞的泄漏。另一方面，大颗粒（&gt; 200纳米; 例如，灰尘或Si碎屑）上的SiN膜可能会导致液体电池的厚度增加，从而导致低的成像造影或低空间分辨率和甚至可能导致的SiN窗破裂。重要的是，盐或其他化学品的残余物可能会影响以不希望的方式的实验的结果。因此，至关重要的是，样品制备和保持器组件的不同步骤仔细和在一个干净，无尘环境中进行。 液体CE的厚度LL确定可实现的分辨率，以及所获得的图像17的对比。该厚度可通过位于两个硅微芯片中的一个间隔物来调整。根据样品的尺寸，液体的细胞的不同的厚度就能够实现。对于金纳米粒子的研究，有可能使用小间隔物（200-500纳米），而整个真核细胞中需要较大的可达5微米的隔离物。液晶单元的厚度是通过从液体小区和周边真空之间的压力差造成的的SiN膜的窗户的膨出进一步影响。这种影响变得具有较大的SiN膜的窗口更加明显。因此，为了最大限度地减少液晶单元的厚度，建议使用小的SiN膜的窗户。在情况下，它是很难找到两个小窗口之间的重叠，它们可以在使用不同的碱微芯片的交叉配置进行组装。其他配置LARG伊利防止胀和由被柱子支撑19单片微型芯片18或膜窗，但那些表现出劣势就装样的。之一的当前技术中最具挑战性的方面是缺乏在液体厚度精确控制的。通常情况下，该液体是比什么是从所用的间隔物的尺寸预期，如在这里所示厚得多。几个研究小组使用封闭液室4，20，21，22;这些系统具有关于空间分辨率的一些优点，作为液体厚度可通过诱导在液体中气泡减少。可替代地，在SiN窗口可以被迫以折叠，导致较薄的液体层。第三，其他窗户更薄的外壳存在（ 例如，石墨烯）23，也导致薄得多液体比什么是可能的，在此协议中描述的系统。然而，这是不可能的那些系统中流动的液体。 作为任何高分辨率显微技术，一些实验方面必须加以考虑。最重要的方面是与液体或样品的电子束的相互作用。除了辐射损伤，这限制了对许多固体样品24中的可实现的空间分辨率，所述液体样品也通过电子束产生的辐解产物15，25的影响。由于这些产品可能会影响实验，细心数据解释和实验设计是必不可少的26。显微镜设置应根据特定研究的目标来选择。 ADF STEM是在较大的液体单元的厚度高原子序数（Z）的成像纳米颗粒更强大，WHI透射电镜乐给出了低Z材料更好的对比度和通常更快，但需要更薄的液体层3。代替使用ADF检测器的，则明场（BF）检测器有时被用于图像的液晶单元中，由于高炉干为在厚层27成像低Z材料是有利的。随着液体电池的厚度，则需要更多的电流。然而，这也增加了辐解产物的浓度和增加辐射伤害。还应当指出的是，对比反转在ADF检测器被观察到很厚的液体（&gt; 10微米为水）。 液体条件通过从显微镜移除保持器和交换两样品和液体我们的实验之间改变。除了改变盐的浓度，这是很容易可以通过在不同的液体流动（ 例如，一个可以改变液体的其它性质为了设置一个特定的pH 16使用缓冲溶液或可能引入的有机溶液或其它添加剂）。另外，也可以改变液体，而保持器在显微镜通过微流体系统流动的液体仍然插入。然而，在这种情况下，它是未知的，此时指向试样的变化的液体中。也值得注意的是，微芯片支承电极是可用的，所以纳米级电化学实验可进行28。 研究的对象不限于金纳米粒子在水，但可使用上述的协议，包括二氧化硅，氧化钛和聚合物进行研究的各种试样。如果对象的运动太快，收购内的图像中捕捉到，可通过一个数量级，通过使用50％甘油和50％水的混合物中来降低粘度。 从上述的点，具有许多优点，可能性，也存在缺点将变得显而易见。当与液相STEM工作，要考虑的最重要的缺点是:1）任何实验是由与整个试样（被观察物，液体，和在SiN膜）的电子束的动态相互作用的影响; 2）样品处理是繁琐的，并且它往往是难以实现的薄液体层，因为样品或微芯片包含一些微米尺寸的颗粒; 3）液体厚度通常从由间隔设置的预定厚度基本上不同;和4）的空间分辨率和对比度强烈取决于液体厚度和下观察和液体中的对象的变化密度之间的差异。 目前，为对象的液体的显微镜用纳米空间分辨率存在充足的方法。非晶冰电子显微镜是一种强大的技术29，但所涉及的实验步骤细腻，不是所有的实验允许样品在冰的制备，和时间分辨实验是不可能的。 X射线显微镜30，31原则上可以使用的，但它有一个有限的空间分辨率，而不是在实验室广泛使用。在液体原子力显微镜已经建立，但是一个表面技术仅32，33，34，35。光学显微镜未显示出足够的空间分辨率。目前，在液体电子显微镜，似乎在液体纳米级物体和过程的直接镜检最强大的技术。 液相TEM和STEM尚未常规分析技术，但仍处于发展阶段。的参数考虑到的数量是相当大的，而且它是ofteÑ难以再现的实验结果。此外，量化数据难以获得，因为所研究的影响与存在的作为电子束的结果进程交织在一起。这里描述的协议的目的是规范实验方案，从而占实验的所有相关方面的基础。我们希望，该协议将导致这一新兴领域的试点工作更好的重复性。

常规扫描透射电子显微镜（STEM）是由适合于分析的标本，特别适合于在高真空放置的干燥和固体样品的范围的限制。然而，许多科学和技术问题涉及纳米级材料和在液体环境中的流程。完全嵌入在液体样品现在可以用STEM使用涉及在透射电子显微镜（TEM）和STEM 1的试样夹持器组装的微流体室中的概念进行了研究。这种新开发的技术已经变得越来越流行，因为它提供了新的认识的各种研究课题重要过程，包括生长，溶解，和纳米颗粒2，3，4，5的聚集方法，6。不仅金属，也biominerals 7和生物系统可以研究8，9，10，11。样品装载和图像获取用于液相干比为干燥样品的干不同并涉及需要专门的培训的协议。 微流体系统包括两个硅微芯片支承氮化硅（SiN）膜的窗户的透明，在200千电子伏的能量12的电子束的（ 见图1A）。尺寸和这些芯片的处理细节可以在别处找到12，13。样品通常含有纳米级物体。在本文中，我们观察到黄金纳米粒子（纳米金）。该金纳米粒子在顶部窗口（相对于向下行进的电子束）固定或浮动在理趣ID。在STEM纳米级的空间分辨率是通过扫描电子束在金纳米颗粒和收集用环形暗场（ADF）的检测器9发送的散射电子获得。两个微芯片被放置在一个小槽在液体流动的TEM保持器1（支架为STEM操作和TEM但作为TEM的保持器被称作）的前端。一项所述的微芯片的包含一个间隔件，以便在微芯片之间形成的液体隔室。在两个微芯片的两侧的O形环提供液体隔室13（ 见图1B）的真空密封。 本文的目的是展示样和数据采集的基本步骤，让有兴趣的用户可能会发现很容易进入这个新兴的新技术。从特定的公司提供的系统被使用，但该协议也适用于其他公司的系统。该技术是比传统的TEM和STEM和一定数量的实际方面较复杂，必须与流体储存器系统13工作时被考虑。最重要的是要确保液体隔室是否正确组装，从而提供了薄液层和真空密封。因此，为了干净地工作，以防止液体流动的TEM支架的制备和组装过程中的粉尘的形成是非常重要的。具体地，O形圈和两个硅微芯片需要从所有污染。灰尘对微芯片的一个甚至小颗粒可能会严重增加组装的电池，其可以防止一个有用的空间分辨率的实现的厚度。真空密封是很重要的，这样没有污染或损坏将在实验后的电子显微镜离开了。本协议描述的加载过程和一些必要的测试。电子显微镜的操作很简单，BUt将其要求相比，固体样品的显微一些额外的步骤。随着液体的厚度，更多的电子被吸收并通过液体散射;液体厚度的测量是必要的。最后，协议说明如何图像并如何动态过程进行研究。 <img alt="图1" src="/files/ftp_upload/54943/54943fig1.jpg" /> 图1:冷却液流量为细胞扫描透射电子显微镜（STEM）。 （A）中组装的液晶单元的示意图。用氮化硅（SiN）膜Windows中两个硅微芯片位于两个O形环之间。由于液体被封闭在SiN膜之间，从而从电子显微镜的真空分开。聚焦电子束扫描过的样本。对比度是由散射电子获得的。金粒子（AuNPs）被固定在所述的SiN膜的液体内，但也可以移动的液体。 2微芯片与O形环的叠层的（B）的示意性侧视图的横截面。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54943/54943fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

<table><tbody><tr><td>Binocular light microscope</td><td>Leica</td><td>M60 CMO</td><td></td></tr><tr><td>Scanning transmission electron mircoscope with spherical aberration corrector</td><td>JEOL</td><td>ARM200F</td><td></td></tr><tr><td>Liquid flow TEM specimen holder</td><td>DENS Solutions</td><td>Ocean</td><td></td></tr><tr><td>Microfluidic syringe pump</td><td>Harvard Scientific</td><td>PicoPlus</td><td></td></tr><tr><td>Plasma cleaner</td><td>Gatan</td><td>Solarus950</td><td></td></tr><tr><td>&lt;strong&gt;Chemicals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Acetone, Rotisolv&amp;nbsp; Plus for HPLC</td><td>Sigma-Aldrich</td><td>7328.2</td><td></td></tr><tr><td>Water, chromasolv Plus for HPLC</td><td>Sigma-Aldrich</td><td>34877-2.5L</td><td></td></tr><tr><td>Ethanol, Rotisolv HPLC grade</td><td>Carl Roth</td><td>P076.2</td><td></td></tr><tr><td>Gold colloid citrate stabilized, diameter 30 nm</td><td>British-Biocell</td><td>EM.GC20</td><td></td></tr><tr><td>&lt;strong&gt;Materials&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Base silicon microchips with silicon nitride membranes of 50 nm thickness and dimensions of 20 &amp;micro;m x 0.40 mm</td><td>DENS Solutions</td><td>for Ocean system</td><td></td></tr><tr><td>Spacer silicon microchips with silicon nitride membranes of 50 nm thickness, dimensions of 20 &amp;micro;m x 0.40 mm, and spacer thickness of 200 nm</td><td>DENS Solutions</td><td>for Ocean system</td><td></td></tr><tr><td>Microfluidic peek tubing</td><td>Upchurch Scientific</td><td>1570</td><td></td></tr><tr><td>Plastic Replaceable tips Tweezers</td><td></td><td></td><td></td></tr><tr><td>(Anti-Magnetic Anti-Acid Stainless Steel body with ESD PVDF (SV) tips)</td><td>ideal-tek</td><td>2ASVR.SA</td><td></td></tr><tr><td>Teflon coated bent steel tweezers (EMS SA with &quot;PTFE&quot; Coating)</td><td>Electron Microscopy Sciences</td><td>78322-7Te</td><td></td></tr><tr><td>Teflon coated broad beak steel tweezers (EMS 2A &quot;PTFE&quot; Coating)</td><td>Electron Microscopy Sciences</td><td>78322-2ATe</td><td></td></tr><tr><td>Hamilton syringe, 1 mL, gastight (Model 1001 TLLX SYR)</td><td>Hamilton</td><td>81323</td><td></td></tr><tr><td>Clean room tissue Sontara Micropure AP (224 x 224mm)</td><td>DuPont</td><td>Sontara MicroPure</td><td></td></tr></tbody></table>

studying dynamic processes of nano-sized objects in liquid using scanning transmission electron microscopy

完全嵌入在液体样品可在使用中用于透射电子显微镜（TEM）和干样品架组装的微流体腔室的扫描透射电子显微镜（STEM）纳米级的空间分辨率进行研究。微流体系统包括两个硅微芯片支撑薄氮化硅（SiN）膜的窗口。本文介绍样品加载和数据采集的基本步骤。最重要的是要确保液体隔室是否正确组装，从而提供了薄液层和真空密封。该协议还包括若干以确保正确装配样品加载期间执行必要的试验。一旦样品在电子显微镜装载，液体厚度需要被测定。不正确装配可能会导致过厚的液体，而太细的液体可以指示不存在液体，当形成的气泡等。最后，协议解释如何图像并如何动态过程进行研究。含有金纳米粒子样品都在纯水和在盐水中进行成像。

液体流动的TEM支架被用来研究金纳米粒子在液体中的行为。金纳米粒子被稳定地固定在SiN膜在纯水中，并使用液相STEM（ 图6）与纳米级的分辨率进行成像。在强烈散射黄金获得良好的对比度。用于干试样测得的荧光屏上的电流密度为20 PA /平方厘米，而其量达8 PA /平方厘米与插入的液体流动的TEM持有者。使用公式1， 叔 水 = 2.4±0.5微米，比基于为200nm的隔离物厚度是预期的大得多。然而，厚度不为金纳米粒子的纳米空间分辨率成像太大。液体厚度由于鼓出的微芯片的氮化硅膜，非平坦的，和碎片驻留在微芯片比由间隔设置的200纳米厚。 16中它们的形状，虽然反应性辐解产物（例如- 水溶液 ，H•，H +，OH•）从电子束的与交互始发水可能氧化单金原子，导致纳米金15的形状的变化。然而，当液体流动系统被用来在第二个实验中引入的氯离子，所述金纳米粒子的稳定性发生变化。氯离子能够稳定在tetrachloroaureat的形式氧化金原子，AUCL 4 - 。 图7示出，在一个STEM成像时间推移系列缓慢溶解的金纳米粒子，类似的结果报告早期的16。对于所使用的电子剂量率，花〜300秒以溶解30纳米大小的金纳米粒子。 金纳米粒子的水:运动<html> R IN第三个实验中（ 图8）进行了研究。之前的实验中，将液体流动的TEM保持器以除去盐的任何痕迹进行清洗。从第一个实验的不同，使用一个替代的样品制备的方法来实现的纳米金来在SiN膜14的较弱的附着。在该实验中，金纳米粒子溶液置于在硅微芯片，并在液体流动的TEM保持器组装而不让溶液变干。以这种方式，所述金纳米颗粒容易从在SiN膜在成像在所使用的剂量率分离。一些金纳米粒子的移动离开的视图成批量溶液的字段路程，而其余的金纳米粒子在靠近在SiN窗口保持在视野内。观察这些金纳米粒子的运动，最终他们团聚。一段时间后，这些聚集体也从在SiN膜分离并移出的视场并进入溶液中。 ntent"FO:保together.within页="1"&gt; <img alt="图6" src="/files/ftp_upload/54943/54943fig6.jpg" /> 图6:顶部有一个纯净水层的金纳米粒子30纳米直径在扫描透射电子显微镜（STEM）显微图。示出的图像是原始图像的选定区域。图像大小为1,024×1,024像素，像素停留时间为19微秒，像素尺寸为0.73纳米，并且放大倍率为400,000X。电子剂量为7.1因而×10 4 E - /纳米2。在荧光屏上测得的电流密度为8 PA /厘米2，使该液体的厚度经计算为等于2.4微米。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54943/54943fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图7" src="/files/ftp_upload/54943/54943fig7.jpg" /> 图7:时间推移系列在盐水金纳米粒子STEM显微照片。 （AD）的图片从在30秒的间隔时间推移系列STEM图像的提取。该金纳米粒子在液体逐渐溶解作为氯离子的存在下的结果。的像素停留时间为2微秒，时间经过一系列的帧时间为1.75秒，像素尺寸为0.44纳米，并且放大倍率为500,000。每幅图像的电子剂量为1.2×10 4 E - /纳米2。液体厚度为2.4微米。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54943/54943fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图8" src="/files/ftp_upload/54943/54943fig8.jpg" /> 图8:STEM金纳米粒子在纯净水移动显微照片。 （A）的SiN膜与金纳米粒子，其中一些是用箭头选中。 （ 二 ）运动轨迹所选择的金纳米粒子（见A）。一些金纳米粒子从视野中成像时搬走。剩下的金纳米粒子沿着氮化硅薄膜横向移动，并开始凝聚。在到达临界簇大小，它们从膜调度和从view.The像素停留时间字段搬开为1微秒，则帧时间为0.52秒，像素尺寸为1.8纳米，并且放大倍率为120,000X。每幅图像的电子剂量为3.5×10 2ë - /纳米2和液体厚度为2.4μm。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54943/54943fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy at JoVE.com

Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

这个协议描述一种用于扫描金纳米粒子的透射电子显微镜中的水，如用于纳米动态过程的观察液流试样保持器的操作。

使用扫描透射电子显微镜研究纳米大小的物体的动态过程中的液体

Samples fully embedded in liquid can be studied at a nanoscale spatial resolution with Scanning Transmission Electron Microscopy (STEM) using a ...

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Universidad San Sebastian

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JoVE Journal

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12.6K 次观看.  INM-Leibniz Institute for New Materials. 这个协议描述一种用于扫描金纳米粒子的透射电子显微镜中的水，如用于纳米动态过程的观察液流试样保持器的操作。 

视频: Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

Revealing Dynamic Processes of Materials in Liquids Using Liquid Cell Transmission Electron Microscopy

The recent development for in situ transmission electron microscopy, which allows imaging through liquids with high spatial resolution, has attracted significant interests across the research fields of materials science, physics, chemistry and biology. The key enabling technology is a liquid cell. We fabricate liquid cells with thin viewing windows through a sequential microfabrication process, including silicon nitride membrane deposition, photolithographic patterning, wafer etching, cell bonding, etc. A liquid cell with the dimensions of a regular TEM grid can fit in any standard TEM sample holder. About 100 nanoliters reaction solution is loaded into the reservoirs and about 30 picoliters liquid is drawn into the viewing windows by capillary force. Subsequently, the cell is sealed and loaded into a microscope for in situ imaging. Inside the TEM, the electron beam goes through the thin liquid layer sandwiched between two silicon nitride membranes. Dynamic processes of nanoparticles in liquids, such as nucleation and growth of nanocrystals, diffusion and assembly of nanoparticles, etc., have been imaged in real time with sub-nanometer resolution. We have also applied this method to other research areas, e.g., imaging proteins in water. Liquid cell TEM is poised to play a major role in revealing dynamic processes of materials in their working environments. It may also bring high impact in the study of biological processes in their native environment.

We have developed a self-contained liquid cell, which allows imaging through liquids using a transmission electron microscope. Dynamic processes of nanoparticles in liquids can be revealed in real time with sub-nanometer resolution.

在液体中使用液体池透射电子显微镜揭示材料的动态过程

The recent development for in situ transmission electron microscopy, which allows imaging through liquids with high spatial resolution, has attracted ...

科研

工程学

Quantitative and Qualitative Examination of Particle-particle Interactions Using Colloidal Probe Nanoscopy

Colloidal Probe Nanoscopy (CPN), the study of the nano-scale interactive forces between a specifically prepared colloidal probe and any chosen substrate using the Atomic Force Microscope (AFM), can provide key insights into physical interactions present within colloidal systems. Colloidal systems are widely existent in several applications including, pharmaceuticals, foods, paints, paper, soil and minerals, detergents, printing and much more.1-3 Furthermore, colloids can exist in many states such as emulsions, foams and suspensions. Using colloidal probe nanoscopy one can obtain key information on the adhesive properties, binding energies and even gain insight into the physical stability and coagulation kinetics of the colloids present within. Additionally, colloidal probe nanoscopy can be used with biological cells to aid in drug discovery and formulation development. In this paper we describe a method for conducting colloidal probe nanoscopy, discuss key factors that are important to consider during the measurement, and show that both quantitative and qualitative data that can be obtained from such measurements.

Colloidal probe nanoscopy can be used within a variety of fields to gain insight into the physical stability and coagulation kinetics of colloidal systems and aid in drug discovery and formulation sciences using biological systems. The method described within provides a quantitative and qualitative means to study such systems.

粒子粒子相互作用采用胶体探针纳米显微定量和定性检查

Colloidal Probe Nanoscopy (CPN), the study of the nano-scale interactive forces between a specifically prepared colloidal probe and any chosen substrate ...

化学

In situ TEM of Biological Assemblies in Liquid

Researchers regularly use Transmission Electron Microscopes (TEMs) to examine biological entities and to assess new materials. Here, we describe an additional application for these instruments- viewing viral assemblies in a liquid environment. This exciting and novel method of visualizing biological structures utilizes a recently developed microfluidic-based specimen holder. Our video article demonstrates how to assemble and use a microfluidic holder to image liquid specimens within a TEM. In particular, we use simian rotavirus double-layered particles (DLPs) as our model system. We also describe steps to coat the surface of the liquid chamber with affinity biofilms that tether DLPs to the viewing window. This permits us to image assemblies in a manner that is suitable for 3D structure determination. Thus, we present a first glimpse of subviral particles in a native liquid environment.

In situ TEM of Biological Assemblies in Liquid

Here we describe a procedure to image viral complexes in liquid at nanometer resolution using a transmission electron microscope.

就地 液体生物组件的 TEM

Researchers regularly use Transmission Electron Microscopes (TEMs) to examine biological entities and to assess new materials. Here, we describe an ...

生物工程

Sub-nanometer Resolution Imaging with Amplitude-modulation Atomic Force Microscopy in Liquid

Atomic force microscopy (AFM) has become a well-established technique for nanoscale imaging of samples in air and in liquid. Recent studies have shown that when operated in amplitude-modulation (tapping) mode, atomic or molecular-level resolution images can be achieved over a wide range of soft and hard samples in liquid. In these situations, small oscillation amplitudes (SAM-AFM) enhance the resolution by exploiting the solvated liquid at the surface of the sample. Although the technique has been successfully applied across fields as diverse as materials science, biology and biophysics and surface chemistry, obtaining high-resolution images in liquid can still remain challenging for novice users. This is partly due to the large number of variables to control and optimize such as the choice of cantilever, the sample preparation, and the correct manipulation of the imaging parameters. Here, we present a protocol for achieving high-resolution images of hard and soft samples in fluid using SAM-AFM on a commercial instrument. Our goal is to provide a step-by-step practical guide to achieving high-resolution images, including the cleaning and preparation of the apparatus and the sample, the choice of cantilever and optimization of the imaging parameters. For each step, we explain the scientific rationale behind our choices to facilitate the adaptation of the methodology to every user's specific system.

We present a method for achieving sub-nanometer resolution images with amplitude-modulation (tapping mode) atomic force microscopy in liquid. The method is demonstrated on commercial atomic force microscopes. We explain the rationale behind our choices of parameters and suggest strategies for resolution optimization.

亚纳米级分辨率成像与调幅原子力显微镜在液体

Atomic force microscopy (AFM) has become a well-established technique for nanoscale imaging of samples in air and in liquid. Recent studies have shown ...

Liquid-cell Transmission Electron Microscopy for Tracking Self-assembly of Nanoparticles

Drying a nanoparticle dispersion is a versatile way to create self-assembled structures of nanoparticles, but the mechanism of this process is not fully understood. We have traced the trajectories of individual nanoparticles using liquid-cell transmission electron microscopy (TEM) to investigate the mechanism of the assembly process. Herein, we present the protocols used for liquid-cell TEM studies of the self-assembly mechanism. First, we introduce the detailed synthetic protocols used to produce uniformly sized platinum and lead selenide nanoparticles. Next, we present the microfabrication processes used to produce liquid cells with silicon nitride or silicon windows and then describe the loading and imaging procedures of the liquid-cell TEM technique. Several notes are included to provide helpful tips for the entire process, including how to manage the fragile cell windows. The individual motions of nanoparticles tracked by liquid-cell TEM revealed that changes in the solvent boundaries caused by evaporation affected the self-assembly process of nanoparticles. The solvent boundaries drove nanoparticles to primarily form amorphous aggregates, followed by flattening of the aggregates to produce a 2-dimensional (2D) self-assembled structure. These behaviors are also observed for different nanoparticle types and different liquid-cell compositions.

Here we introduce experimental protocols for the real-time observation of a self-assembly process using liquid-cell transmission electron microscopy.

用于跟踪纳米粒子自组装的液体细胞透射电子显微镜

Drying a nanoparticle dispersion is a versatile way to create self-assembled structures of nanoparticles, but the mechanism of this process is not fully ...

All-electronic Nanosecond-resolved Scanning Tunneling Microscopy: Facilitating the Investigation of Single Dopant Charge Dynamics

The miniaturization of semiconductor devices to scales where small numbers of dopants can control device properties requires the development of new techniques capable of characterizing their dynamics. Investigating single dopants requires sub-nanometer spatial resolution, which motivates the use of scanning tunneling microscopy (STM). However, conventional STM is limited to millisecond temporal resolution. Several methods have been developed to overcome this shortcoming, including all-electronic time-resolved STM, which is used in this study to examine dopant dynamics in silicon with nanosecond resolution. The methods presented here are widely accessible and allow for local measurement of a wide variety of dynamics at the atomic scale. A novel time-resolved scanning tunneling spectroscopy technique is presented and used to efficiently search for dynamics.

We demonstrate an all-electronic method to observe nanosecond-resolved charge dynamics of dopant atoms in silicon with a scanning tunneling microscope.

全电子纳秒分辨扫描隧道显微镜: 促进单一掺杂剂电荷动力学的研究

The miniaturization of semiconductor devices to scales where small numbers of dopants can control device properties requires the development of new ...

Using Graphene Liquid Cell Transmission Electron Microscopy to Study in Situ Nanocrystal Etching

Graphene liquid cell electron microscopy provides the ability to observe nanoscale chemical transformations and dynamics as the reactions are occurring in liquid environments. This manuscript describes the process for making graphene liquid cells through the example of graphene liquid cell transmission electron microscopy (TEM) experiments of gold nanocrystal etching. The protocol for making graphene liquid cells involves coating gold, holey-carbon TEM grids with chemical vapor deposition graphene and then using those graphene-coated grids to encapsulate liquid between two graphene surfaces. These pockets of liquid, with the nanomaterial of interest, are imaged in the electron microscope to see the dynamics of the nanoscale process, in this case the oxidative etching of gold nanorods. By controlling the electron beam dose rate, which modulates the etching species in the liquid cell, the underlying mechanisms of how atoms are removed from nanocrystals to form different facets and shapes can be better understood. Graphene liquid cell TEM has the advantages of high spatial resolution, compatibility with traditional TEM holders, and low start-up costs for research groups. Current limitations include delicate sample preparation, lack of flow capability, and reliance on electron beam-generated radiolysis products to induce reactions. With further development and control, graphene liquid cell may become a ubiquitous technique in nanomaterials and biology, and is already being used to study mechanisms governing growth, etching, and self-assembly processes of nanomaterials in liquid on the single particle level.

Using Graphene Liquid Cell Transmission Electron Microscopy to Study in Situ Nanocrystal Etching

Graphene liquid cell electron microscopy can be used to observe nanocrystal dynamics in a liquid environment with greater spatial resolution than other liquid cell electron microscopy techniques. Etching premade nanocrystals and following their shape using graphene liquid cell Transmission Electron Microscopy can yield important mechanistic information about nanoparticle transformations.

用石墨烯液细胞透射电镜研究原位纳米晶蚀刻

Graphene liquid cell electron microscopy provides the ability to observe nanoscale chemical transformations and dynamics as the reactions are occurring in ...

Nanoscale Characterization of Liquid-Solid Interfaces by Coupling Cryo-Focused Ion Beam Milling with Scanning Electron Microscopy and Spectroscopy

Physical and chemical processes at solid-liquid interfaces play a crucial role in many natural and technological phenomena, including catalysis, solar energy and fuel generation, and electrochemical energy storage. Nanoscale characterization of such interfaces has recently been achieved using cryogenic electron microscopy, thereby providing a new path to advancing our fundamental understanding of interface processes.
This contribution provides a practical guide to mapping the structure and chemistry of solid-liquid interfaces in materials and devices using an integrated cryogenic electron microscopy approach. In this approach, we pair cryogenic sample preparation which allows stabilization of solid-liquid interfaces with cryogenic focused ion beam (cryo-FIB) milling to create cross-sections through these complex buried structures. Cryogenic scanning electron microscopy (cryo-SEM) techniques performed in a dual-beam FIB/SEM enable direct imaging as well as chemical mapping at the nanoscale. We discuss practical challenges, strategies to overcome them, as well as protocols for obtaining optimal results. While we focus in our discussion on interfaces in energy storage devices, the methods outlined are broadly applicable to a range of fields where solid-liquid interface play a key role.

Cryogenic Focused Ion Beam (FIB) and Scanning Electron Microscopy (SEM) techniques can provide key insights into the chemistry and morphology of intact solid-liquid interfaces. Methods for preparing high quality Energy Dispersive X-ray (EDX) spectroscopic maps of such interfaces are detailed, with a focus on energy storage devices.

通过冷冻聚焦离子束铣削与扫描电子显微镜和光谱学耦合，对液固界面进行纳米级表征

Physical and chemical processes at solid-liquid interfaces play a crucial role in many natural and technological phenomena, including catalysis, solar ...

Studying the Effects of Temperature on the Nucleation and Growth of Nanoparticles by Liquid-Cell Transmission Electron Microscopy

Temperature control is a recent development that provides an additional degree of freedom to study nanochemistry by liquid cell transmission electron microscopy. In this paper, we describe how to prepare an in situ heating experiment for studying the effect of temperature on the formation of gold nanoparticles driven by radiolysis in water. The protocol of the experiment is fairly simple involving a special liquid cell with uniform heating capabilities up to 100 &#176;C, a liquid-cell TEM holder with flow capabilities and an integrated interface for controlling the temperature. We show that the nucleation and growth mechanisms of gold nanoparticles are drastically impacted by the temperature in liquid cell. Using STEM imaging and nanodiffraction, the evolution of the density, size, shape and atomic structure of the growing nanoparticles are revealed in real time. Automated image processing algorithms are exploited to extract useful quantitative data from video sequences, such as the nucleation and growth rates of nanoparticles. This approach provides new inputs for understanding the complex physico-chemical processes at play during the liquid-phase synthesis of nanomaterials.

Temperature control during liquid-phase electron microscopy experiments opens up new perspectives of studying the dynamic of nanoparticles in liquid environments mimicking their formation or application media. Using recently developed heating liquid cells, we directly observed the influence of temperature on the nucleation and growth processes of gold nanoparticles in water.

研究温度对液细胞传输电子显微镜对纳米粒子核化和生长的影响

Temperature control is a recent development that provides an additional degree of freedom to study nanochemistry by liquid cell transmission electron ...

Light-Induced In Situ Transmission Electron Microscopy for Observation of the Liquid-Soft Matter Interaction

The current protocol describes the modifications of the transmission electron microscope (TEM) setup for in situ light-induced observations. A glass optical fiber inserted into the electron column above the objective lens polepiece, and a laser, an adjustable light source, was used to fabricate the device. After the illuminator has been calibrated using an external measuring system, it allows one to adjust the intensity of the lighting to the needs of the observed process. This lighting system was utilized to image antimicrobial photodynamic therapy phenomena, which are currently the subject of intense research. The sample was prepared by spotting a suspension of bacteria on a carbon, graphene, or silicon nitride substrate, blotting the excess solution, spotting the photosensitizer solution, blotting the excess liquid again, and then assembling the liquid cell with a second substrate or graphene film. The process of the imaging experiment itself includes choosing the right place for observation with the use of low magnification and a minimum dose of electrons, and then cyclical activation of the light source to capture subsequent images at specified intervals with the minimum amount of electrons necessary. The electron dose of each exposure and the time and intensity of lighting used need to be carefully recorded due to the complexity of the observed phenomena as, at the same time, the process is both light- and electron-driven. After the actual experiment is performed, additional control observations must be made, in which the same doses of electrons are used but without additional light influence and smaller doses of electrons are used for higher doses of light. This makes it possible to distinguish light-induced microstructural effects from those caused by electrons in both the fields of life and materials science.

Light-Induced In Situ Transmission Electron Microscopy for Observation of the Liquid-Soft Matter Interaction

The present protocol describes transmission electron microscope (TEM) modifications with a light illumination system, the fabrication of liquid cells, and in situ TEM observations of light-induced interactions between bacterial cells and a photosensitizer. The sample preparation methods, electron beam damage, and imaging are also discussed.

光诱导 原 位透射电子显微镜观察液-软物质相互作用

The current protocol describes the modifications of the transmission electron microscope (TEM) setup for in situ light-induced observations. A glass ...

使用扫描透射电子显微镜研究纳米大小的物体的动态过程中的液体

摘要

探索更多视频

此视频中的章节

在液体中使用液体池透射电子显微镜揭示材料的动态过程

粒子粒子相互作用采用胶体探针纳米显微定量和定性检查

就地液体生物组件的 TEM

亚纳米级分辨率成像与调幅原子力显微镜在液体

用于跟踪纳米粒子自组装的液体细胞透射电子显微镜

全电子纳秒分辨扫描隧道显微镜: 促进单一掺杂剂电荷动力学的研究

用石墨烯液细胞透射电镜研究原位纳米晶蚀刻

通过冷冻聚焦离子束铣削与扫描电子显微镜和光谱学耦合，对液固界面进行纳米级表征

研究温度对液细胞传输电子显微镜对纳米粒子核化和生长的影响

光诱导原位透射电子显微镜观察液-软物质相互作用

使用扫描透射电子显微镜研究纳米大小的物体的动态过程中的液体

摘要

探索更多视频

此视频中的章节

在液体中使用液体池透射电子显微镜揭示材料的动态过程

粒子粒子相互作用采用胶体探针纳米显微定量和定性检查

就地 液体生物组件的 TEM

亚纳米级分辨率成像与调幅原子力显微镜在液体

用于跟踪纳米粒子自组装的液体细胞透射电子显微镜

全电子纳秒分辨扫描隧道显微镜: 促进单一掺杂剂电荷动力学的研究

用石墨烯液细胞透射电镜研究原位纳米晶蚀刻

通过冷冻聚焦离子束铣削与扫描电子显微镜和光谱学耦合，对液固界面进行纳米级表征

研究温度对液细胞传输电子显微镜对纳米粒子核化和生长的影响

光诱导 原 位透射电子显微镜观察液-软物质相互作用

就地液体生物组件的 TEM

光诱导原位透射电子显微镜观察液-软物质相互作用