The authors have no acknowledgements.

注意:PTCDA可刺激皮肤或眼睛，因此应谨慎使用合适的手套进行处理。请参考相应的安全手册。低温液体可以产生类似于热烧伤皮肤上的效果或会造成长时间暴露冻伤。处理低温液体时，一定要佩戴防护眼镜及适当的低温手套。通过低温液体形成的气体是非常冷，通常比空气重，可以积累在地板置换空气附近。当没有足够的空气或氧气，窒息死亡，可能会发生。请参考相应的安全手册。 1.样品制备<ol><li> PTCDA对Ag的沉积（111） 注意:PTCDA的单层（ML）的范围应为10-30％，和集中在大，紧凑的岛屿（ 图6）之间。这种情况是理想的，而具有尖端preparati足够的清洁金属表面做处理实验上。 <ol><li>用溅射和退火周期14的标准程序在沉积前清洁的Ag（111）晶。 <ol><li>溅射用Ar +离子晶体15分钟。使用氩压力在1×10 -5毫巴，0.8千电子伏的离子能量，并在室温（RT）的晶体。 </li><li>退火在530℃下将样品15-30分钟。 </li></ol></li><li>使用PTCDA K-细胞在室温14的银矿床PTCDA 0.1-0.3 ML（111）的样品。 注意:没有沉积参数给出的，因为沉积条件可从设置不同设置。 </li><li>沉积之后，刷新样品至200℃进行2分钟，以提高PTCDA岛屿的排序和解吸可能的污染。 </li><li>或者，通过检查PTCDA对Ag（111）15衍射图案与检查LEED的沉积。 </li><li>使用特定的设置，步骤将样品转移到SPM。通常情况下，使用manipulatoř能够内特高压和可能的手动操作摆动棒线性转移。 </li></ol></li><li> 检查在SPM样品制备。 <ol><li>样品转移后等到了SPM的温度接近其基准温度（在这里:5 K）。在所描述的设置的持续时间为约1小时的样品转移期间冷却至液氮温度。 </li><li>使用具体的设置，程序到尖端接近到表面（在恒定电流模式），直至出现隧道电流。 </li><li>选择高压放大器的偏移电压，例如，美Z = 0。这将在整个纸的标准设置，如果没有另外指明。 </li><li>检查样品的制备制备恒流STM图像（设定点:I = 0.1 nA的，偏压V B = -0.35 V施加到样品，电流放大器的增益为1×10 9 V / A）的表面上。给定的参数促进最低unoccupie的成像PTCDA d的分子轨道（LUMO）。这有助于确定选择用于操纵（插图图6）的分子的羧基氧原子的位置。 </li><li>制备的前端直到STM图像类似于图6，对于在尖部-表面的间隔距离对应于当前成像设置点或由7-10埃移动至前端（从稳定点5-6Ⅴ的例子使用脉冲）向干净的Ag（111）面，同时施加V b = 0.1 V至样品。崩溃的前端更深在双尖端的情况下。不要使用接近PTCDA岛脉冲！ </li></ol></li></ol> 2.设置AFM操作与TFS <ol><li>设置为FM-AFM系统特定的PLL参数，使得ΔF检测是可能与可接受的噪音的条件和检测速度（ 例如，0.1-0.5 Hz的噪声中的ΔF在约0.2-0.4埃的所述TFS振幅于7赫兹带宽）。 </li><li&gt;检查Q因数（由软件自动计算），并判断所述TFS的f 0的。 <ol><li>缩回从表面的前端到SPM控制器能够达到（ 例如，通过设置V Z = -10 V这在此处描述的设置的情况下，大约180埃缩回从表面的尖端）的最大距离。 </li><li>记录使用SPM软件的共振曲线（TFS振幅与在恒定的TFS激励振幅驱动频率）。 </li><li>读出的谐振频率f 0为最大，在频率轴上的谐振曲线的位置。 Q因子是由从谐振峰的宽度的软件计算。所描述的设置的Q因子之间50,000-70,000（ 图7）而变化。 </li></ol></li><li>在一个干净的Ag（111）面面积的尖端定位和校准以下文献所述TFS振荡幅度。 16。 </li></ol> 3.集成Ø˚FMCS在SPM设置<ol><li>组装和根据从制造商得到的手动校准的MCS。校准包括设置坐标系MCS的原点。 </li><li>下面就到系统手动开关，并将其添加在MCS软件跟踪的对象。 </li><li>检查跟踪由检测量移动和之后由MCS软件所显示的位置正常工作。 </li><li>测试RVS并通过从窗口发送测试命令RVS软件之间的连接（参见图4）。 </li><li>测试的MCS，RVS和TipControl之间的连接。 <ol><li>检查是否在V 点¯x - ，Vÿ - ，RVS诉ž-voltages被设置为0 V，如果需要重新设置。 <ol><li>缩回从表面（2.2.1）的尖端。 </li><li>按"重置所有"按钮，在软件窗口重置v 点¯x - ，Vÿ - ，Vž-voltages在RVS的输出。 </li><li>一个接近角尖回用FB环表面关闭（1.2.2）。 </li></ol></li><li>使用SPM软件的具体设置功能在干净的Ag（111）表面的尖端位置。 </li><li>检查的x，y，z轴复选框在软件窗口。这激活沿所有三个空间轴的尖端位置的手工控制模式。 </li><li>按"开始"，在软件窗口。 </li><li>确保在V 点¯x - ，Vÿ - ，-voltages通过RVS产生vž正确地向沿着每个轴的运动作出反应。而沿z轴移动（垂直于表面），监视的FB回路，试图补偿从RVS施加vŽ -电压的反应。 </li><li>在软件窗口中按"暂停"。 </li><li>在软件窗口中按"复位所有"。 </li></ol></li></ol> 4. HMD集成在SPM设置<ol><li>确保HMD连接和所有必要的DRIVERS根据来自制造商的手册进行安装。 </li><li>启动VRinterface并确保它正确呈现模型表面，吸附分子和末端（参见图5）。 </li><li>对准在HMD看到与MCS的坐标轴的三维虚拟现实场景的坐标系的方向。 </li><li>戴上HMD。如果需要，重新定位HMD在你的头上，而执行以下步骤要么观看VR场景或实验室环境，键盘和电脑显示器。 </li><li>测试I和ΔF信号的实时数据传输从SPM电子， 例如，通过改变SPM软件内的隧道电流设定点。 <ol><li>检查X，Y，软件窗口的z轴的复选框。 </li><li>拿起至，然后按软件窗口中的"开始"按钮。 </li><li>移动TO并检查是否代表尖端领域是虚拟三维场景内移动正确。 </li><li>保持手握住稳住，直到软件窗口的"暂停"按钮被按下。 </li><li>收起了TO。 </li><li>按"复位所有"软件窗口的按钮。 </li></ol></li></ol> 5.准备SPM单PTCDA分子的操纵<ol><li>设置在恒定电流模式下的STM与便于PTCDA LUMO对比度的参数，从而允许一个，以确定分子取向（设定点:I = 0.1 nA的，偏置电压V B =施加到样品-0.35伏，电流放大器增益1×10 9 V / A）。 </li><li>确保尖端是操作做好准备。 <ol><li>图像PTCDA。在SPM软件，扫描输入的参数（区域进行扫描（如300×300 2），对于反馈环路设定点:I = 0.1 nA和V B = -0.35 V，扫描速度= 150纳米/秒），然后按在SPM软件中的"开始"按钮。的图像的分辨率必须是类似于图6。 </li><li>确保当尖端从隧道接触移动以从表面（&gt; 100埃）的距离大，发生Δf是不超过5-7赫兹大得多。 </li><li>在情况下，任何的上述条件未能重复在干净的Ag（111）面（1.2.5）的前端制备。 </li></ol></li><li>找到适于操纵的表面积。 <ol><li>使用的SPM软件查找类似于图6，其中包含一个PTCDA岛和干净的Ag（111）面的某些区域所示的一个的区域。如果需要，用干净的区域，重塑不同的操作尝试之间的一角。 </li><li>选择PTCDA岛内的分子进行操纵和记录了详细的STM图像（ 例如 ，50×50埃2）， 如图6中选择。 -从下拉菜单中选择"SetXYOffset顶"，选择区域用于通过详细的图像点击较大的概述图片。 注:有没有特别的标准，因为一个小岛（约3分子远离边缘）中的所有分子可以被认为是相等的操作。不应该有"猛料"上可见或旁边分子。这样污垢会在图像中产生不规则的对比度。 </li></ol></li><li>测试该尖端结合到PTCDA分子的能力。 <ol><li>以上PTCDA的使用SPM软件的具体的设置功能的两个羧酸氧原子（标于图6）中的一个的尖端的位置。选择"SetXYOffset - 顶"，然后单击相应的形象。 </li><li>记录在其中的前端由3-5埃和I（z）的垂直移动朝向表面被使用SPM软件的具体的设置的函数记录的光谱。 <ol><li>设置一个恒定偏压V B（ 例如，6毫伏）并限定尖端高度的斜坡接近和缩回从表面的前端（ 例如 ，4埃;接近，然后再返回）。然后在SPM软件点击按钮"VERT。MANIP"，并选择最近录制的STM图像，在垂直的操作应执行上的位置。 </li></ol></li><li>检查，如果所记录的I（z）的表现出的尖端，并在尖锐（所记录的频谱以外Z分辨率）形式的分子之间的接触形成的电流I（z）的增加。通常情况下，接触0.5-3埃通过纵向回缩尖起重足够强（ 见图8）。 <ol><li>如果I（z）的曲线上没有明显的急剧形成接触尝试以下之一: <ol><li>稍微改变尖端的横向位置，并重复方法步骤。 </li><li>做一个温和尖端成形（1.2.5），并尝试直到象在图8中注册中所示的接触的行为的分子再次接触。 </li></ol></li></ol></li></ol></li><li>一直等到piezØ蠕变消失（约2-4小时）。 注:漂移的量HCM中确定接触点的稳定性，从而可以执行与在同一分子连续操作而不重新扫描区域多久。 <ol><li>沿着通过比较选择操作区域两个详细STM图像，记录的时间间隔， 例如 5分钟的x，y方向测试蠕变。等到漂移是在两个连续图像之间的小于0.5埃。 </li><li>通过记录由FB回路在1分钟内应用U Z（t）和计算出的漂移速率在z方向测试蠕变。杜Z（T）/ dt的应为约0.2埃/小时。 </li></ol></li></ol> 6.准备手控操作（HCM） <ol><li>确保所有相关的程序正在运行，而且连接的设备之间的数据传输正常工作:MCS，TipControl，RVS，VRinterface，HMD和SPM电子产品。 </li><li>确保倾向¼在HMD的坐标系的n被与MCS的坐标轴对准。 </li><li>对齐如图VRinterface视觉辅助与真实分子的取向在实验被操纵该分子的图像。 <ol><li>对准HMD沿其坐标系和定位它，这样的观点是参照分子之上。通过按键盘上的相应按钮顺时针旋转或逆时针中VRinterface参照分子对准在SPM软件成像分子。 </li></ol></li><li>检查是否在V 点¯x - ，Vÿ - ，RVS诉ž-voltages被设置为0 V，如果需要（3.5.1）进行重置。 </li><li>重新扫描选择操作与恒流模式的STM的PTCDA分子。 </li><li>过选择用于使用SPM软件的相应的功能的操作的羧基氧原子的尖端的位置。如在5.4确定使用正确的接触点。 </li> &lt;LI&gt;激活PLL并设置幅度控制模式。设置振荡振幅尽可能低（ 例如，0.2-0.4埃），但足够高以使得ΔF检测是可能与可接受的噪音的条件和检测速度（见2.1）。 </li><li>打开FB循环。在SPM软件参数窗口积分值输入0。 </li><li>设置结偏压到几毫伏的SPM软件参数窗口。输入0.007至7毫伏应用到表面上。 </li><li>设置当前放大器的增益，以1×10 7中的SPM的软件参数窗口V / A。 </li></ol> 7.使用HCM为PTCDA受控操作<ol><li>戴上HMD，并采取TO。如果需要的话，重新定位在HMD上的使用者的头部而执行以下步骤，要么查看VR场景或实验室环境，键盘和计算机显示器。 </li><li>置所记录的轨迹的颜色对比来登录（I（X，Y，Z））在VRinterface按相应的button。 </li><li>标志着3D虚拟场景的接触点。这个"锚"有助于很容易地找到的联系人使用HCM无需重置RVS进一步操纵的企图。 <ol><li>只有通过检查提示软件相应的复选框激活沿Z轴手动控制，同时保持X，Y复选框选中。 </li><li>移动至向下边看我（0,0，z）和ΔF（0,0，z）的实时信号在虚拟场景。停止移动于:当I（0,0，z）和ΔF（0,0，z）的信号表示同时大幅上升，接触地层的签名（参见图8）。 </li><li>按对应的按钮启动VRinterface轨迹记录，并开始移动为UP。 </li><li>在VRinterface尽快停止轨迹记录通过按下相应的按钮，分子和小费破裂之间的接触。签名是I（X，Y，Z）和ΔF（X，Y，Z）的信号的同时急剧下降。 </li><li>按 ＆＃34;在尖端软件暂停"按钮即可停用手控。 </li></ol></li><li>通过在软件检查的x，y，z轴复选框激活沿所有空间轴的尖端运动的手控，并按下在尖端软件的"开始"按钮。 </li><li>万一的接触形成的点从"锚定"在虚拟场景的操作（由于漂移或尖端顶点的任何变化）之后的有偏差修正的前端位置，并在需要的前端的状态。 <ol><li>通过移动一边看在虚拟场景中的白色球体的移动开始手控制之前移动至前端回到其初始位置。 </li><li>在尖端软件按"暂停"按钮即可停用手控。 </li><li>按"重置所有"按钮，在提示软件复位v 点¯x - ，Vÿ - ，RVS诉ž-voltages为0 V. </li><li>以促进LUMO c中的参数设置STM早在恒流模式ontrast的PTCDA（参见1.2.4）。 </li><li>重新扫描选用操纵分子，并使用SPM软件的具体设置，功能尖端超过所选择的羧基氧原子（5.4判断）正确的位置进行定位。如果需要，准备尖在附近的位置（&lt;300之遥），以减少残留的压电蠕变。 </li><li>重新启动协议步骤7.1。 </li></ol></li><li>试着找一个成功的升降轨道，其中接触分子从表面的轨迹的结束完全不沾边。 <ol><li>接近，其中"锚"通过移动TO而以下表示在虚拟场景中的当前前端位置的球的运动表现出形成尖部 - 分子的接触点。只要接触形成记录开始在VRinterface一个新的轨迹。 </li><li>通过移动相应地拉在适于提升的方向（ 图10）的分子。如果检测尖端分子接触的破裂，停止记录轨迹。返回到接触点，启动上形成接触轨迹的记录和执行不同的操作。 </li><li>切换到ΔF（X，Y，Z）所记录的轨迹通过在从表面更大的距离按压VRinterface相应的按钮的颜色对比（在与电流放大器增益约为7埃10 7 V / A），因为I（X， Y，Z）信号衰减从表面快速离开。这里ΔF（X，Y，Z）成为分子的存在的唯一指示器（参见图1）。当分子尖端接触丢失，ΔF（X，Y，Z）跳跃（接近）零，甚至不上接近表面为1-3埃了变化。 </li><li>如果尖端分子接触是Z&gt; 10的静态稳定，注意在ΔF（X，Y，Z）签名的地方显示了从表面拉离分子顺利过渡到零。这是为successf签名分子的微升解除（参照图1）。 </li><li>测试，如果分子完全从表面分离，并挂在前端。 <ol><li>移动到最多检查ΔF（X，Y，Z）上进一步尖端缩回保持在零。 </li><li>移动下来，以检查是否ΔF（X，Y，Z）在接近地面1-3一种超越地方检测成功吊装签名的高度增加。 </li></ol></li></ol></li><li>沉积抬起分子到一个干净的Ag（111）的表面积。 <ol><li>吊装成功后，将高达收回尖端从表面的额外10-20。这减少了与表面的抬起分子的任何相互作用。 </li><li>在尖端软件按"暂停"按钮来修复当前的尖端位置，并停用手动控制。 </li><li>如果没有接通FB循环，使用SPM软件的具体设置，功能定位在清洁银尖（111）表面的一些DISTANCE（如 50-100），从那里提取分子岛之遥。选择"SetXYOffset - 顶"，然后单击相应的形象。 </li><li>设置电流放大器的增益为1×10 9 V / A。 </li><li>检查尖端的软件，然后按仅Z复选框尖端软件的"开始"按钮。 </li><li>移动向表面接近，直到一个可衡量的（Z）出现我。 </li><li>在尖端软件按"暂停"按钮即可停用手控。 </li><li>逐步增加V B通过，直到有在I和ΔF同时跳这表明分子降至在SPM软件使用鼠标控制的滑块（最大V B≈0.5伏，在较高的V B分子可能会损坏）的表面上。如果分子不能被重新沉积，尖端具有待清洁进一步的实验， 例如 ，通过电压脉冲（1.2.5）。 </li><li>扫描区域恒流模式（1.2.4），并检查该分子是否确实沉积背面到表面上。 </li></ol></li></ol>

<ol>
	<li>Barth, J. V., Costantini, G., Kern, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+atomic+and+molecular+nanostructures+at+surfaces.">Engineering atomic and molecular nanostructures at surfaces.</a> Nature. 437, 671-679 (2005).</li><li>Otero, R., Rosei, F., Besenbacher, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scanning+tunneling+microscopy+manipulation+of+complex+organic+molecules+on+solid+surfaces.">Scanning tunneling microscopy manipulation of complex organic molecules on solid surfaces.</a> Annu. Rev. Phys. Chem. 57, 497-525 (2006).</li><li>Urgel, J. I., Ecija, D., Auw&#228;rter, W., Barth, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+Manipulation+of+Gadolinium+Coordinated+Supramolecules+by+Low-Temperature+Scanning+Tunneling+Microscopy.">Controlled Manipulation of Gadolinium Coordinated Supramolecules by Low-Temperature Scanning Tunneling Microscopy.</a> Nano Lett. 14, 1369-1373 (2014).</li><li>Fournier, N., Wagner, C., Weiss, C., Temirov, R., Tautz, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Force-controlled+lifting+of+molecular+wires.">Force-controlled lifting of molecular wires.</a> Phys. Rev. B. 84, 035435 (2011).</li><li>Wagner, C., Fournier, N., Tautz, F. S., Temirov, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+the+Binding+Energies+of+the+Organic-Metal+Perylene-Tetracarboxylic-Dianhydride/Au(111)+Bonds+by+Molecular+Manipulation+Using+an+Atomic+Force+Microscope.">Measurement of the Binding Energies of the Organic-Metal Perylene-Tetracarboxylic-Dianhydride/Au(111) Bonds by Molecular Manipulation Using an Atomic Force Microscope.</a> Phys. Rev. Lett. 109 (7), 076102 (2012).</li><li>Wagner, C., 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=Non-additivity+of+molecule-surface+van+der+Waals+potentials+from+force+measurements.">Non-additivity of molecule-surface van der Waals potentials from force measurements.</a> Nat. Commun. 5, 5568 (2014).</li><li>Green, M. F. B., 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=Patterning+a+hydrogen-bonded+molecular+monolayer+with+a+hand-controlled+scanning+probe+microscope.">Patterning a hydrogen-bonded molecular monolayer with a hand-controlled scanning probe microscope.</a> Beilstein Journal of Nanotechnology. 5, 1926-1932 (2014).</li><li>Leinen, P., 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=Virtual+reality+visual+feedback+for+hand-controlled+scanning+probe+microscopy+manipulation+of+single+molecules.">Virtual reality visual feedback for hand-controlled scanning probe microscopy manipulation of single molecules.</a> Beilstein J. Nanotechnol. 6, 2148-2153 (2015).</li><li>Wagner, C., 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=Scanning+Quantum+Dot+Microscopy.">Scanning Quantum Dot Microscopy.</a> Phys. Rev. Lett. 115 (2), 026101 (2015).</li><li>Mura, 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=Experimental+and+theoretical+analysis+of+H-bonded+supramolecular+assemblies+of+PTCDA+molecules.">Experimental and theoretical analysis of H-bonded supramolecular assemblies of PTCDA molecules.</a> Phys. Rev. B. 81 (19), 195412 (2010).</li><li>Besocke, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+easily+operable+scanning+tunneling+microscope.">An easily operable scanning tunneling microscope.</a> Surf. Sci. Lett. (1-2), 145-153 (1987).</li><li>Giessibl, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+atomic+force+microscopy.">Advances in atomic force microscopy.</a> Rev. Mod. Phys. 75 (3), 949-983 (2003).</li><li>Albrecht, T. R., Gru&#776;tter, P., Horne, D., Rugar, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frequency+modulation+detection+using+high-Q+cantilevers+for+enhanced+force+microscope+sensitivity.">Frequency modulation detection using high-Q cantilevers for enhanced force microscope sensitivity.</a> J. Appl. Phys. 69 (2), 668-673 (1991).</li><li>Temirov, R., Lassise, A., Anders, F. B., Tautz, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kondo+effect+by+controlled+cleavage+of+a+single-molecule+contact.">Kondo effect by controlled cleavage of a single-molecule contact.</a> Nanotechnology. 19 (6), 065401 (2008).</li><li>Gl&#246;ckler, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+ordered+structures+and+submolecular+scanning+tunnelling+microscopy+contrast+of+PTCDA+and+DM-PBDCI+monolayers+on+Ag(111)+and+Ag(110).">Highly ordered structures and submolecular scanning tunnelling microscopy contrast of PTCDA and DM-PBDCI monolayers on Ag(111) and Ag(110).</a> Surf. Sci. 405 (1), 1-20 (1998).</li><li>Simon, G. H., Heyde, M., Rust, H. -. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recipes+for+cantilever+parameter+determination+in+dynamic+force+spectroscopy:+spring+constant+and+amplitude.">Recipes for cantilever parameter determination in dynamic force spectroscopy: spring constant and amplitude.</a> Nanotechnology. 18 (25), 255503 (2007).</li><li>Rohlfing, M., Temirov, R., Tautz, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+structure+and+scanning+tunneling+data+of+a+prototype+organic-inorganic+interface+PTCDA+on+Ag+(111).">Adsorption structure and scanning tunneling data of a prototype organic-inorganic interface PTCDA on Ag (111).</a> Phys. Rev. B. 76 (11), 115421 (2007).</li><li>Guthold, 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=Controlled+Manipulation+of+Molecular+Samples+with+the+nanoManipulator.">Controlled Manipulation of Molecular Samples with the nanoManipulator.</a> IEEE/ASME Trans. Mechatronics. 5 (2), 189-198 (2000).</li></ol>

The authors have nothing to disclose and no competing financial interests.

像其他基于SPM的方法，在本文中描述的分子操作实验还依赖于对SPM针尖的性质在一定程度上。尖端顶点结构（其不能被完全控制）决定的前端分子键的强度。因此尖部 - 分子接触的强度可能有很大的不同，因此有时可能太低。因此，协议中我们提到的尖端品质和尖端治疗程序的一些基本的测试。然而，更严重的尖端治疗可能在某些情况下需要达到满意的操纵的结果。 <p clas...

低温非接触原子间力/扫描隧道显微镜（LT NC-AFM / STM，在以下简单地称为SPM）是选择用于单个原子或分子1的原子精确操纵的工具- 3。基于SPM的操作一般仅限于两个方面，由一系列突然的，往往随机操纵事件（跳跃）的。这基本上限制在过程中的控制。由单个化学键在一个良好定义的原子的位置接触有问题的分子导致了可以克服这些限制4的方法- 9。在其整个操作的接触分子被连接到SPM针尖使得由尖端的适当的位移在所有三个维度移动的分子成为可能。这为在三维空间中进行的各种复杂的操作过程的可能性。但是接触操作可能是喜通过与表面和/或其他分子在其周围，其可以创建力量大到足以破裂尖端分子接触操纵分子的相互作用ndered。因此，SPM尖端的特定三维轨迹可能会或可能不会导致一个成功的操纵事件。因此，一个问题是如何定义导致当尖端分子键具有有限的强度的情况下，操作的成功完成协议，而与它的环境的操纵分子的相互作用是不先验充分表征。 在这里，这个问题是最直观的方式走近想象。实验者被允许简单地通过移动他们手中7控制SPM尖端的位移。这是通过将SPM连接到商用运动捕捉系统实现的，部分规格，其中提供如下。 "手控操纵"（HCM）的优点是在T他实验者的快速尝试不同的操作轨迹，并从他们的失败或成功学习的能力。 的HCM设置已被用来进行验证的原理的实验，其中一个字（"利希"）的苝-3,4,9,10-四羧酸二酐（PTCDA）分子上的Ag的封闭层钢印（ 111），除去48分子，一个接一个，与HCM 7。起重从表面切割其结合的分子在单层10的分子间氢键的分子。通常本分子间键的总强度超过前端的最外原子和通过其中分子接触（ 见图 1）PTCDA的羧酸氧原子之间的单个化学键的强度。可能导致尖端分子接触的破裂和操纵的企图的以下故障。实验者的任务从而determi网元，打破了抵制分子间键顺序地而不是同时，使得施加到尖部 - 分子接触的总力的尖端轨迹永远不会超过其强度。 虽然所希望的轨迹原则上可以模拟，由于系统的尺寸和复杂性所涉及的必要的模拟将采取过于大量的时间。在对比的是，使用HCM有可能后40分钟，除去第一分子。朝向实验结束提取已经采取了这证实了学习过程的有效性的时间少得多。此外，HCM方法的精度和通用性被证明在反向操作的动作时从邻近位置提取的分子被用来关闭从单层的误除去另一个分子的后留下的空隙。 动作捕捉的方式，而被快速和直观，是限于尖轨迹数据的生成。为新的分子操作的协议进一步系统发展它能够实时查看尖端轨迹数据以及分析先前生成的数据是同样重要的。因此，HCM设置的功能是通过将虚拟现实护目镜，其允许实验者看在前端的轨迹被电流（I）增加了三维虚拟场景中绘制的数据和频率偏移基本上增强（ΔF）值测通过实时8 SPM的（见下文）。除此之外，虚拟现实场景示出了操纵分子充当视觉尺度参考的模型。因此，HCM设置由虚拟现实界面称赞适于操纵轨迹空间的系统的映射和的有前途的操纵协议逐次提炼。除了该系统还便于d的知识转移ifferent实验。下面的段落给出了设置的说明和它的一些规范，是相关的操作实验。 该实验在超高真空（UHV）以1×10 -10毫巴用由制备室的市售的SPM和分析室的基本压力进行。该准备容室是配备有:氩离子源用于样品溅射，通过操纵样品转移（允许加热和样品的冷却），低能量电子衍射（LEED），含PTCDA粉末定制克努森细胞（K-细胞）升华纯化。分析室配备有:LN 2浴低温恒温器具有12升的体积和46小时的保持时间，液氦浴低温恒温器（5升，72小时），Besocke 11甲虫型SPM配备有音叉传感器12（ TFS）由石英音叉的带有电连接PTIR尖端（对于STM操作），它被切割和通过聚焦离子束（FIB）（ 图 2）削尖。 <img alt="图1" src="/files/ftp_upload/54506/54506fig2.jpg" /> 图2. 音叉传感器（a）中附PTIR尖端商业音叉传感器的图像。 （ 二 ）切断与FIB的PTIR顶端尖端的SEM图像。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54506/54506fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 原子力显微镜在频率调制（FM），模式13，其中的TFS是在共振（f 0的≈31080赫兹）与抖动压电激励操作。振荡音叉的压电信号通过锁相环（PLL），其保持所述TFS的振荡常数的幅度和跟踪的i变化放大并且用于TS共振频率，ΔF= F - f 0的，即作用于尖端力的梯度起源。如在图 3中所示的SPM针尖位置由电压，Y，Z压电体（压电常数以5K控制（U X，U Y，U z）的应用到一组X轴的:X = 15，Y = 16，Z = 6 A / V）。在U X，U Y，Už-voltages（±10 V，20位分辨率）在SPM电子输出生成的。它们是由具有±200伏的最大输出电压的高电压（HV）放大器进一步放大<img alt="图1" src="/files/ftp_upload/54506/54506fig3.jpg" /> 图 3. HCM设置的 原理图 。的（跟踪对象）的位置已安装在其表面上是由运动捕捉系统（MCS）的两个红外摄像机跟踪多个（红外线）IR来源。 TipControl所以ftware获得从MCS坐标（x，Y，Z），并将其传递到远程电压源（RVS），它产生一组电压（V X，V Y，V z）的被求和的电压（U 点¯x ，U Y，U z）除以SPM电子的SPM针尖位置的控制生产。所添加的电压通过高电压（HV）放大器，并且进一步施加到SPM尖端的压电定位系统。该设置允许尖端定位手动控制时SPM反馈（FB）循环是开放的。前端的（X，Y，Z）位置以及I（X，Y，Z）和ΔF（X，Y，Z）被传递到该绘制它在由操作者看到的3D虚拟场景的VRinterface软件戴着头戴式显示器（HMD）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54506/54506fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 该SPM尖端和之间流动的隧道电流表面由一个跨阻抗放大器测量与从1×10 3的范围内，以1×10 9 V / A的可变增益（带宽增益1×10 9 V / A为1千赫兹）。放大器的输出被馈送到STM反馈（FB）循环来调节恒流扫描模式下的表面上方的针尖高度。结（与所述TFS振荡关闭）的稳定性是1-3时。所述TFS的压电振荡信号进行放大在两个阶段:（1）前置放大器固定至LN 2罩（获得1×10 8 V / A，带宽20千赫），和（2）从1可变增益外部电压放大器×10 1至5×10 4和1兆赫的带宽。 对于HCM实验，SPM设置被扩展为:运动捕捉系统（MCS），远程控制多路电压源（RVS），加法器和虚拟现实头盔显示器（HMD）。所有除summi列出的设备NG放大器在商业上获得的。 MSC是一个红外（IR）标记的跟踪系统，其允许在100赫兹的速率空间位移毫米的分辨率。该系统由两个红外摄像机，可跟踪对象（TO）和控制软件。在MCS软件通过分析由两个照相机获得其图象得到的x，y，在三维空间中的TO的z坐标。 MCS提供了一个编程库，允许在一个单独的软件程序使用的坐标。 TO的坐标（x 的 ，Y 向 ，Z 向 ）传递给定制开发的软件程序"TipControl"。 图 4显示了图形用户界面的屏幕截图。该软件是由在窗口中的"开始"按钮激活。激活后（τ= 0），软件将所有V 点¯x - ，Vÿ - ，Vž-voltages上RVS（±10 V 16电压范围根据以下表达式位分辨率，每电压步骤50毫秒延迟） <img alt="式（1）" src="/files/ftp_upload/54506/54506eq1.jpg" /> 等 ，其中c X，C Y，C z是该转换5cm至的位移装入SPM尖端1的位移的因素。的因素P X [（T），对Y（T），P Z（T）具有由X轴的状态定义的值，Y，Z复选框在软件窗口中。如果被检查的框，那么相应的P（T）被设置为1所有的P（t）的时刻，当在软件窗口按下"暂停"按钮被设置为0。允许操作者暂时"冻结"前端的位置。在软件窗口中按下"重置所有"按钮，如下设置V 点¯x - ，Vÿ - ，Vž-voltages零返回尖端由SPM软件定义其初始位置。在软件窗口中CA的文本字段"手册命令RVS"<html> N为用于设置任何在V x的- ，Vÿ - ，VŽ-voltages到在±10 V在V x的允许范围内的任何值- ，Vÿ - ，-voltages由RVS相加产生vž到U 点¯x - ，UŸ -通过求和放大器SPM电子üž -输出电压信号（获得1点，带宽为50kHz，输出范围±10 V）。 <img alt="图1" src="/files/ftp_upload/54506/54506fig4.jpg" /> 图4.截图界面窗口的两个指标表现出与MCS和RVS系统连接的状态。复选框用于激活沿选定的空间轴手控。的"启动"按钮，根据图中所示的方案启动的MCS，TipControl和RVS之间的数据流3。按钮"暂停"停止数据流。按钮"全部重设"将所有RVS电压为零。="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54506/54506fig4large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 对实验数据的可视化（尖端轨迹，我，ΔF）头戴式显示器（HMD）被使用。在HMD提供了一个立体视图（拆分高清显示器 - 一半为每只眼睛，1,920×1,080像素为75 Hz）。专用IR照相机跟踪HMD的在3D空间中使用固定的HMD的表面上的IR-LED的位置和方向。该HMD跟踪系统允许操作员通过他们的头一转，或简单地移动自己的身体来改变三维虚拟现实场景里的视图。 定制编写的软件"VRinterface"收集无论从SPM和MCS中的数据，使用OpenGL渲染它在3D场景并用的HMD软件开发工具包（SDK）的帮助下将其显示在HMD。 VRinterface直接从检索实际的x，尖端的y轴，z坐标尖软件（几毫秒的延迟），而I和ΔF信号从SPM电子（延迟≈250毫秒）的输出直接读取。 图 5示出所看到由操作者的HCM期间穿着HMD三维虚拟场景的屏幕快照。里面的3D虚拟场景的顶端尖端呈现白色球体。所记录的前端轨迹的着色反映任一日志（I（X，Y，Z））或ΔF（X，Y，Z）的值。日志之间切换（I（X，Y，Z））或ΔF（X，Y，Z）的色彩模式是通过按下按钮完成。另一个按钮启动记录（和显示）实验尖轨迹数据。当再次按下该按钮停止记录。虚拟场景还示出了被用作操纵期间视觉辅助的静态PTCDA分子。操作者手动对准其取向通过使用键盘上的按钮，以适合表面上的真实分子的取向。 注意事项:由于最前面的tHMD的横移依赖于红外发光二极管，其可以与MCS干扰，因为它也使用IR光来跟踪的位置。因此，要必须有由MCS认可的独特的造型。这有助于MCS到来自TO和那些来自HMD的红外发光二极管来的信号之间的区别。 <img alt="图1" src="/files/ftp_upload/54506/54506fig5.jpg" /> 图5. 在3D虚拟场景第 creenshot HCM期间显示在HMD操作者。一组白色球体形成模型的Ag（111）面。模型表面的取向不一定与样品的取向一致。在PTCDA分子的模型放置在模型表面之上。 C，O，PTCDA的H原子以黑色，红色和白色分别显示。为模型分子的方便方位角取向的目的，可以进行调整，以适应所选择的真实分子的取向进行操纵。前端位置由表示最外层的尖端顶点原子的单个白色球体标记。的实时I（X，Y，Z）和ΔF（X，Y，Z）的数据被显示为下一个放置在提示条的指标。任一日志（I（X，Y，Z））或ΔF（X，Y，Z）对应的轨迹的位置的测量值与当前执行的操作被显示为三维轨迹的颜色表示先前记录为好。该图显示了彩色与log（I（X，Y，Z））的信号轨迹。颜色对比可以登录（I（X，Y，Z））和ΔF（X，Y，Z）由一个按钮，按下模式之间进行切换。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54506/54506fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>LN2</td>
			<td></td>
			<td></td>
			<td>caution: cryogenic liquid</td>
		</tr>
		<tr>
			<td>LHe</td>
			<td></td>
			<td></td>
			<td>caution: cryogenic liquid</td>
		</tr>
		<tr>
			<td>PTCDA</td>
			<td></td>
			<td></td>
			<td>caution: irritating substance</td>
		</tr>
		<tr>
			<td>Knudsen cell (K-cell)</td>
			<td></td>
			<td></td>
			<td>custom</td>
		</tr>
		<tr>
			<td>ErLEED</td>
			<td>Specs</td>
			<td></td>
			<td>used with power supply ErLEED 1000A</td>
		</tr>
		<tr>
			<td>combient LT NC-AFM/STM</td>
			<td>Createc</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>qPlus sensor</td>
			<td>Createc</td>
			<td></td>
			<td>TFS</td>
		</tr>
		<tr>
			<td>preamplifier&#160;</td>
			<td>Createc</td>
			<td></td>
			<td>amplifier for tuning forc signal fixed to LN2 shield (stage 1)</td>
		</tr>
		<tr>
			<td>Low-Noise Voltage Preamplifier</td>
			<td>Standford Research System</td>
			<td>SR560</td>
			<td>external amplifier for tuning forc signal (stage 2)</td>
		</tr>
		<tr>
			<td>Variable Gain Low Noise Current Amplifier</td>
			<td>Femto</td>
			<td>DLPCA-200</td>
			<td>amplifier for tunneling current</td>
		</tr>
		<tr>
			<td>Bonita</td>
			<td>Vicon</td>
			<td>B10, SN: MXBN-0B10-3658</td>
			<td>MCS IR camera</td>
		</tr>
		<tr>
			<td>Apex Interaction Device</td>
			<td>Vicon</td>
			<td>SN: AP0062</td>
			<td>MCS trackable object (TO)</td>
		</tr>
		<tr>
			<td>MX Calibration Wand</td>
			<td>Vicon</td>
			<td></td>
			<td>MCS calibration object</td>
		</tr>
		<tr>
			<td>Tracker</td>
			<td>Vicon</td>
			<td></td>
			<td>MCS software</td>
		</tr>
		<tr>
			<td>BS series voltage supply</td>
			<td>stahl-electronics</td>
			<td>BS 1-4</td>
			<td>RVS</td>
		</tr>
		<tr>
			<td>summing amplifier&#160;</td>
			<td></td>
			<td></td>
			<td>custom, gain 1, based on operational amplifier TL072</td>
		</tr>
		<tr>
			<td>Oculus Rrift Development Kit 2</td>
			<td>Oculus VR</td>
			<td></td>
			<td>HMD</td>
		</tr>
		<tr>
			<td>TipControl</td>
			<td></td>
			<td></td>
			<td>custom-written software</td>
		</tr>
		<tr>
			<td>VRinterface</td>
			<td></td>
			<td></td>
			<td>custom-written software</td>
		</tr>
	</tbody>
</table>

hand controlled manipulation of single molecules via a scanning probe microscope with a 3d virtual reality interface

考虑到有机分子作为未来纳米技术的功能模块，如何安排并以自下而上的方式组装这样的积木的问题仍然是开放的。扫描探针显微镜（SPM）可以是选择的工具;然而，基于SPM的操作，直至最近仅限于二维（2D）。在一个良好定义的位置结合的SPM尖端的分子打开在三维空间中的控制操作的机会。不幸的是，三维操作是与观看和产生的SPM的数据的计算机上的典型2D-范例很大程度上是不相容的。为了直观和高效的操作，因此我们夫妇低温非接触式原子力/扫描隧道显微镜（LT NC-AFM / STM）与运动捕捉系统，充分沉浸式虚拟现实护目镜。这种设置允许"手控操纵"（HCM），其中，所述SPM尖端根据实验者的手的移动而移动，WHILE尖端轨迹以及在三维被可视化的SPM结的响应。 HCM铺平了道路的复杂的操作协议的发展，可能导致在表面上的分子之间起作用纳米相互作用的更好基本理解。在这里，我们描述了设置和实现虚拟现实环境中成功手控分子操作所需的步骤。

注意:此部分显示发表在7,8工作。 HCM应用起重PTCDA /银（111）出层的问题，我们可以通过依次删除单个分子（ 图 9）写的模式。在总共48个分子中除去，其中40可以再沉积到清洁的Ag（111），表明该分子留在操作过程不变。这允许使用HCM通过从不同的位置采取分子和灌装的意外创建空缺（ ?...

Watch this Scientific Journal Video about Hand Controlled Manipulation of Single Molecules via a Scanning Probe Microscope with a 3D Virtual Reality Interface at JoVE.com

Hand Controlled Manipulation of Single Molecules via a Scanning Probe Microscope with a 3D Virtual Reality Interface

We demonstrate the precise manipulation of individual organic molecules on a metal surface with the tip of a scanning probe microscope driven in 3D by the experimenter's hand using a motion capture system and fully immersive virtual reality goggles.

通过手扫描探针显微镜用三维虚拟现实接口控制单分子的操作

Considering organic molecules as the functional building blocks of future nanoscale technology, the question of how to arrange and assemble such building ...

hand-controlled-manipulation-single-molecules-via-scanning-probe

Research

JoVE Journal

Engineering

8.9K 次观看.  Forschungszentrum Jülich. We demonstrate the precise manipulation of individual organic molecules on a metal surface with the tip of a scanning probe microscope driven in 3D by the experimenter's hand using a motion capture system and fully immersive virtual reality goggles.

视频: Hand Controlled Manipulation of Single Molecules via a Scanning Probe Microscope with a 3D Virtual Reality Interface

High-resolution, High-speed, Three-dimensional Video Imaging with Digital Fringe Projection Techniques

Digital fringe projection (DFP) techniques provide dense 3D measurements of dynamically changing surfaces.&#160;Like the human eyes and brain, DFP uses triangulation between matching points in two views of the same scene at different angles to compute depth.&#160;However, unlike a stereo-based method, DFP uses a digital video projector to replace one of the cameras1. The projector rapidly projects a known sinusoidal pattern onto the subject, and the surface of the subject distorts these patterns in the camera&#8217;s field of view. Three distorted patterns (fringe images) from the camera can be used to compute the depth using triangulation.
Unlike other 3D measurement methods, DFP techniques lead to systems that tend to be faster, lower in equipment cost, more flexible, and easier to develop.&#160;DFP systems can also achieve the same measurement resolution as the camera.&#160;For this reason, DFP and other digital structured light techniques have recently been the focus of intense research (as summarized in1-5). Taking advantage of DFP, the graphics processing unit, and optimized algorithms, we have developed a system capable of 30&#160;Hz 3D video data acquisition, reconstruction, and display for over 300,000 measurement points per frame6,7.&#160;Binary defocusing DFP methods can achieve even greater speeds8.
Diverse applications can benefit from DFP techniques. Our collaborators have used our systems for facial function analysis9, facial animation10, cardiac mechanics studies11, and fluid surface measurements, but many other potential applications exist.&#160;This video will teach the fundamentals of DFP techniques and illustrate the design and operation of a binary defocusing DFP system.

This video describes the fundamentals of digital fringe projection techniques, which provide dense 3D measurements of dynamically changing surfaces.&#160;It also demonstrates the design and operation of a high-speed binary defocusing system based on these techniques.

Digital fringe projection (DFP) techniques provide dense 3D measurements of dynamically changing surfaces.&#160;Like the human eyes and brain, DFP uses ...

科研

工程学

Construction of a High Resolution Microscope with Conventional and Holographic Optical Trapping Capabilities

High resolution microscope systems with optical traps allow for precise manipulation of various refractive objects, such as dielectric beads 1 or cellular organelles 2,3, as well as for high spatial and temporal resolution readout of their position relative to the center of the trap. The system described herein has one such "traditional" trap operating at 980 nm. It additionally provides a second optical trapping system that uses a commercially available holographic package to simultaneously create and manipulate complex trapping patterns in the field of view of the microscope 4,5 at a wavelength of 1,064 nm. The combination of the two systems allows for the manipulation of multiple refractive objects at the same time while simultaneously conducting high speed and high resolution measurements of motion and force production at nanometer and piconewton scale.

The system described herein employs a traditional optical trap as well as an independent holographic optical trapping line, capable of creating and manipulating multiple traps. This allows for the creation of complex geometric arrangements of refractive particles while also permitting simultaneous high-speed, high-resolution measurements of the activity of biological enzymes.

高分辨率显微镜与常规和全息光学捕获能力建设

High resolution microscope systems with optical traps allow for precise manipulation of various refractive objects, such as dielectric beads 1 or cellular ...

Scanning-probe Single-electron Capacitance Spectroscopy

The integration of low-temperature scanning-probe techniques and single-electron capacitance spectroscopy represents a powerful tool to study the electronic quantum structure of small systems - including individual atomic dopants in semiconductors. Here we present a capacitance-based method, known as Subsurface Charge Accumulation (SCA) imaging, which is capable of resolving single-electron charging while achieving sufficient spatial resolution to image individual atomic dopants. The use of a capacitance technique enables observation of subsurface features, such as dopants buried many nanometers beneath the surface of a semiconductor material1,2,3. In principle, this technique can be applied to any system to resolve electron motion below an insulating surface.
As in other electric-field-sensitive scanned-probe techniques4, the lateral spatial resolution of the measurement depends in part on the radius of curvature of the probe tip. Using tips with a small radius of curvature can enable spatial resolution of a few tens of nanometers. This fine spatial resolution allows investigations of small numbers (down to one) of subsurface dopants1,2. The charge resolution depends greatly on the sensitivity of the charge detection circuitry; using high electron mobility transistors (HEMT) in such circuits at cryogenic temperatures enables a sensitivity of approximately 0.01 electrons/Hz&frac12; at 0.3 K 5.

Scanning-probe single-electron capacitance spectroscopy facilitates the study of single-electron motion in localized subsurface regions. A sensitive charge-detection circuit is incorporated into a cryogenic scanning probe microscope to investigate small systems of dopant atoms beneath the surface of semiconductor samples.

扫描探针单电子电容谱

The integration of low-temperature scanning-probe techniques and single-electron capacitance spectroscopy represents a powerful tool to study the ...

Single Plane Illumination Module and Micro-capillary Approach for a Wide-field Microscope

A module for light sheet or single plane illumination microscopy (SPIM) is described which is easily adapted to an inverted wide-field microscope and optimized for 3-dimensional cell cultures, e.g., multi-cellular tumor spheroids (MCTS). The SPIM excitation module shapes and deflects the light such that the sample is illuminated by a light sheet perpendicular to the detection path of the microscope. The system is characterized by use of a rectangular capillary for holding (and in an advanced version also by a micro-capillary approach for rotating) the samples, by synchronous adjustment of the illuminating light sheet and the objective lens used for fluorescence detection as well as by adaptation of a microfluidic system for application of fluorescent dyes, pharmaceutical agents or drugs in small quantities. A protocol for working with this system is given, and some technical details are reported. Representative results include (1) measurements of the uptake of a cytostatic drug (doxorubicin) and its partial conversion to a degradation product, (2) redox measurements by use of a genetically encoded glutathione sensor upon addition of an oxidizing agent, and (3) initiation and labeling of cell necrosis upon inhibition of the mitochondrial respiratory chain. Differences and advantages of the present SPIM module in comparison with existing systems are discussed.

A module for single plane illumination microscopy (SPIM) is described which is easily adapted to an inverted wide-field microscope and optimized for 3-dimensional cell cultures. The sample is located within a rectangular capillary, and via a microfluidic system fluorescent dyes, pharmaceutical agents or drugs can be applied in small quantities.

单一平面照明模块和一个宽场显微镜微毛细管法

A module for light sheet or single plane illumination microscopy (SPIM) is described which is easily adapted to an inverted wide-field microscope and ...

Energy Dispersive X-ray Tomography for 3D Elemental Mapping of Individual Nanoparticles

Energy dispersive X-ray spectroscopy within the scanning transmission electron microscope (STEM) provides accurate elemental analysis with high spatial resolution, and is even capable of providing atomically resolved elemental maps. In this technique, a highly focused electron beam is incident upon a thin sample and the energy of emitted X-rays is measured in order to determine the atomic species of material within the beam path. This elementally sensitive spectroscopy technique can be extended to three dimensional tomographic imaging by acquiring multiple spectrum images with the sample tilted along an axis perpendicular to the electron beam direction.
Elemental distributions within single nanoparticles are often important for determining their optical, catalytic and magnetic properties. Techniques such as X-ray tomography and slice and view energy dispersive X-ray mapping in the scanning electron microscope provide elementally sensitive three dimensional imaging but are typically limited to spatial resolutions of &gt; 20 nm. Atom probe tomography provides near atomic resolution but preparing nanoparticle samples for atom probe analysis is often challenging. Thus, elementally sensitive techniques applied within the scanning transmission electron microscope are uniquely placed to study elemental distributions within nanoparticles of dimensions 10-100 nm.
Here, energy dispersive X-ray (EDX) spectroscopy within the STEM is applied to investigate the distribution of elements in single AgAu nanoparticles. The surface segregation of both Ag and Au, at different nanoparticle compositions, has been observed.

The use of energy dispersive X-ray tomography in the scanning transmission electron microscope to characterize elemental distributions within single nanoparticles in three dimensions is described.

能量色散X射线断层摄影术对个人纳米3D元素映射

Energy dispersive X-ray spectroscopy within the scanning transmission electron microscope (STEM) provides accurate elemental analysis with high spatial ...

Comprehensive Characterization of Extended Defects in Semiconductor Materials by a Scanning Electron Microscope

Extended defects such as dislocations and grain boundaries have a strong influence on the performance of microelectronic devices and on other applications of semiconductor materials. However, it is still under debate how the defect structure determines the band structure, and therefore, the recombination behavior of electron-hole pairs responsible for the optical and electrical properties of the extended defects. The present paper is a survey of procedures for the spatially resolved investigation of structural and of physical properties of extended defects in semiconductor materials with a scanning electron microscope (SEM). Representative examples are given for crystalline silicon. The luminescence behavior of extended defects can be investigated by cathodoluminescence (CL) measurements. They are particularly valuable because spectrally and spatially resolved information can be obtained simultaneously. For silicon, with an indirect electronic band structure, CL measurements should be carried out at low temperatures down to 5 K due to the low fraction of radiative recombination processes in comparison to non-radiative transitions at room temperature. For the study of the electrical properties of extended defects, the electron beam induced current (EBIC) technique can be applied. The EBIC image reflects the local distribution of defects due to the increased charge-carrier recombination in their vicinity. The procedure for EBIC investigations is described for measurements at room temperature and at low temperatures. Internal strain fields arising from extended defects can be determined quantitatively by cross-correlation electron backscatter diffraction (ccEBSD). This method is challenging because of the necessary preparation of the sample surface and because of the quality of the diffraction patterns which are recorded during the mapping of the sample. The spatial resolution of the three experimental techniques is compared.

The optical, electrical, and structural properties of dislocations and of grain boundaries in semiconductor materials can be determined by experiments performed in a scanning electron microscope. Electron microscopy has been used to investigate cathodoluminescence, electron beam induced current, and diffraction of backscattered electrons.

通过扫描电子显微镜在半导体材料的扩展缺陷的全面表征

Extended defects such as dislocations and grain boundaries have a strong influence on the performance of microelectronic devices and on other applications ...

Scanning SQUID Study of Vortex Manipulation by Local Contact

Local, deterministic manipulation of individual vortices in type 2 superconductors is challenging. The ability to control the position of individual vortices is necessary in order to study how vortices interact with each other, with the lattice, and with other magnetic objects. Here, we present a protocol for vortex manipulation in thin superconducting films by local contact, without applying current or magnetic field. Vortices are imaged using a scanning superconducting quantum interference device (SQUID), and vertical stress is applied to the sample by pushing the tip of a silicon chip into the sample, using a piezoelectric element. Vortices are moved by tapping the sample or sweeping it with the silicon tip. Our method allows for effective manipulation of individual vortices, without damaging the film or affecting its topography. We demonstrate how vortices were relocated to distances of up to 0.8 mm. The vortices remained stable at their new location up to five days. With this method, we can control vortices and move them to form complex configurations. This technique for vortex manipulation could also be implemented in applications such as vortex based logic devices.

We present a protocol for manipulation of individual vortices in thin superconducting films, using local mechanical contact. The method does not include applying current, magnetic field or additional fabrication steps.

由当地接触式扫描涡操纵SQUID研究

Local, deterministic manipulation of individual vortices in type 2 superconductors is challenging. The ability to control the position of individual ...

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 ...

Single-Digit Nanometer Electron-Beam Lithography with an Aberration-Corrected Scanning Transmission Electron Microscope

We demonstrate extension of electron-beam lithography using conventional resists and pattern transfer processes to single-digit nanometer dimensions by employing an aberration-corrected scanning transmission electron microscope as the exposure tool. Here, we present results of single-digit nanometer patterning of two widely used electron-beam resists: poly (methyl methacrylate) and hydrogen silsesquioxane. The method achieves sub-5 nanometer features in poly (methyl methacrylate) and sub-10 nanometer resolution in hydrogen silsesquioxane. High-fidelity transfer of these patterns into target materials of choice can be performed using metal lift-off, plasma etch, and resist infiltration with organometallics.

We use an aberration-corrected scanning transmission electron microscope to define single-digit nanometer patterns in two widely-used electron-beam resists: poly (methyl methacrylate) and hydrogen silsesquioxane. Resist patterns can be replicated in target materials of choice with single-digit nanometer fidelity using liftoff, plasma etching, and resist infiltration by organometallics.

用畸变校正扫描透射电镜进行单数字纳米电子束光刻

We demonstrate extension of electron-beam lithography using conventional resists and pattern transfer processes to single-digit nanometer dimensions by ...

Visualizing Uniaxial-strain Manipulation of Antiferromagnetic Domains in Fe1+YTe Using a Spin-polarized Scanning Tunneling Microscope

The quest to understand correlated electronic systems has pushed the frontiers of experimental measurements toward the development of new experimental techniques and methodologies. Here we use a novel home-built uniaxial-strain device integrated into our variable temperature scanning tunneling microscope that enables us to controllably manipulate in-plane uniaxial strain in samples and probe their electronic response at the atomic scale. Using scanning tunneling microscopy (STM) with spin-polarization techniques, we visualize antiferromagnetic (AFM) domains and their atomic structure in Fe1+yTe samples, the parent compound of iron-based superconductors, and demonstrate how these domains respond to applied uniaxial strain. We observe the bidirectional AFM domains in the unstrained sample, with an average domain size of ~50-150 nm, to transition into a single unidirectional domain under applied uniaxial strain. The findings presented here open a new direction to utilize a valuable tuning parameter in STM, as well as other spectroscopic techniques, both for tuning the electronic properties as for inducing symmetry breaking in quantum material systems.

Visualizing Uniaxial-strain Manipulation of Antiferromagnetic Domains in Fe1+YTe Using a Spin-polarized Scanning Tunneling Microscope

Using uniaxial strain combined with spin-polarized scanning tunneling microscopy, we visualize and manipulate the antiferromagnetic domain structure of Fe1+yTe, the parent compound of iron-based superconductors.

利用自旋偏振扫描隧道显微镜对 fe 1+yTe 中的反铁磁领域进行单轴应变操作

The quest to understand correlated electronic systems has pushed the frontiers of experimental measurements toward the development of new experimental ...