Name: hyperspectral imaging as a tool to study optical anisotropy in lanthanide-based molecular single crystals
Uploaded: 2020-04-14T14:00:00
Description: Watch this Scientific Journal Video about Hyperspectral Imaging as a Tool to Study Optical Anisotropy in Lanthanide-Based Molecular Single Crystals at JoVE.com

作者感谢渥太华大学化学和生物分子科学系的迪伦·埃鲁拉特先生和穆拉利·穆鲁格苏教授提供[TbEu（bpm）（tfaa）6] 单晶。6E.M.R.、N.R.和E.H.感谢渥太华大学、加拿大创新基金会和加拿大自然科学和工程研究理事会（NSERC）提供的财政支助。

注意：建议在操作成像器时，使用针对任何时候都使用的激发波长特有的安全护目镜。
1. 高光谱显微镜的配置
注：图2a概述了高光谱成像系统，并介绍了成像器的主要组件。成像系统可用于检测样品中的可见或近红外 （NIR） 发射。根据所需的检测（可见或近红外），光通过两个不同的光路（图2e）。不同的光?...

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此处描述的高光谱成像协议提供了一种直接的方法，允许在样品的精确位置获取光谱信息。使用所述设置，空间分辨率（x和y映射）可以达到 0.5 μm，而可见范围内的映射的光谱分辨率可以达到 0.2 nm，NIR 范围的光谱分辨率为 0.6 nm。
为了对单个晶体进行高光谱映射，样品制备遵循一个简单的过程：晶体可以简单地放置在玻璃显微镜幻灯片上，根据需要由盖玻璃覆?...

高光谱成像（HSI）是一种生成空间地图的技术，其中每个x-y坐标包含一个光谱信息，这些信息可以基于任何类型的光谱学，即光致发光、吸收和散射光谱1,1、2、3。,3因此，获得了一组三维数据集（也称为"高光谱立方体"），其中x-y坐标为空间轴，z坐标是分析样本的光谱信息。因此，高光谱立方体包含空间和光谱信息，比传统光谱学对样品进行更详细的光谱研究。,6,7,虽然HSI在遥感领域（如地质学、食品工业4）已有多年所知，但最近它已成为一种用于生物医学应用的纳米材料2、55或探针的特性创新2技术。一般来说，它不限于紫外线/可见/近红外（NIR）领域，而且还可以使用其他辐射源，如X射线扩展，例如，为了描述不同材料9或Terahertz辐射中的元素分布特征，其中HSI用于在生物组织8中进行热传传。此外，光致发光映射与拉曼映射相结合，以探测单层MoS210的光学特性。然而，在光学HSI的应用中，关于基于兰他尼的材料的HSI的例子仍然只有几个例子：11、12、13、14、15、16、17。11,12,13,14,15,16,17例如，我们可以举一个例子：发现组织中的癌症6，分析生物组织中的光穿透深度7，多路复用生物成像3，混合系统中多组分能量转移分析11，以及研究向上转换纳米粒子的聚合诱导光谱特性变化12。显然，HSI的吸引力源于它适合生成关于环境特定发光的知识，同时提供有关探测器的空间和光谱信息。
利用这一强大的技术，本文描述了一种研究异核Tb3+-Eu3+单晶[TbEu（bpm）（tfaa）6] （6图1a）13的光学自给性的协议。13观测到的光相向异性是由于Ln3+离子在不同晶体方向中的分子包装不同（图1b），导致一些晶体表面显示更亮，其他晶体表面显示发光。有人建议，晶体特定表面的发光强度增加与Ln3=[]的晶体方向的更有效能量传输有关。Ln3+ ion 距离最短的13。
在这些结果的激励下，我们建议建立一个详细的方法，通过HSI分析光学各向异性，为更好地了解电离能量转移过程和特定分子排列产生的可调发光特性开辟道路。,19这些结构特性关系被公认为创新光学材料设计的重要方面，包括但不限于纳米和微尺度的波导系统和光磁存储设备，满足对更高效、小型化光学系统20的需求。

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			<td height="21" style="height:21px;width:216px;">Microscope glass slides</td>
			<td style="width:123px;">FisherBrand</td>
			<td style="width:161px;">12-550-15</td>
			<td style="width:167px;">Glass slides used for sample preparation</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Visible and Near Infrared Hyperspectral Confocal Imager</td>
			<td style="width:123px;">PhotonETC</td>
			<td style="width:161px;"></td>
			<td>Microscope used for the analysis, builted according to the user needs, therefore it is no catalog number</td>
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hyperspectral imaging as a tool to study optical anisotropy in lanthanide-based molecular single crystals

在本作品中，我们描述了高光谱成像（HSI）在分析基于发光的蓝硅（Ln3+）分子单晶时的新应用协议。作为具有代表性的例子，我们选择了基于异核Ln的复合物[TbEu（bpm）（tfaa）6] （bpm=2，2'-双丙氨酸，tfaa= =1，1，1-三氟乙酰聚酰亚酸盐）的单晶，在紫外线激发下表现出明亮的可见发射。6HSI是一种新兴技术，将发光结构的二维空间成像与所获取图像的每个像素的光谱信息相结合。具体来说，[Tb-Eu] 复合体单晶上的HSI提供了局部光谱信息，揭示了所研究晶体不同点的发光强度变化。这些变化归因于晶体中的光学自给性，这是由于Ln3+离子在晶体结构的每个方向中不同的分子包装造成的。本文描述的HSI是这种技术适合分子材料光谱空间研究的一个例子。然而，重要的是，该协议可以很容易地扩展到其他类型的发光材料（如微米大小的分子晶体，无机微粒，生物组织中的纳米粒子，或标记的细胞等），为更深入地研究结构与性质的关系开辟了许多可能性。最终，此类调查将为从生物成像到技术应用（如波导或光电器件）等各种应用的高级材料工程提供可供利用的知识。

为了说明高光谱显微镜在基于Ln的分子单晶（即[TbEu（bpm）（tfaa）6+，图1a）上采集数据的配置，图2显示了系统的概述以及光学立方体在设置中的正确位置。6图 3显示了包含 HSI 采集期间使用的菜单的 PHySpec 软件的屏幕截图。图 4和图 5更详细地显示了显微镜阶段，包括包含要分?...

Watch this Scientific Journal Video about Hyperspectral Imaging as a Tool to Study Optical Anisotropy in Lanthanide-Based Molecular Single Crystals at JoVE.com

Hyperspectral Imaging as a Tool to Study Optical Anisotropy in Lanthanide-Based Molecular Single Crystals

在这里，我们提出了一个协议，以获得发光高光谱成像数据，并使用高光谱成像系统分析基于兰他尼的单晶体的光自给特征。

高光谱成像作为研究基于兰塔尼的分子单晶光学自给性的工具

In this work, we describe a protocol for a novel application of hyperspectral imaging (HSI) in the analysis of luminescent lanthanide (Ln3+)-based ...

高光谱成像允许在样品中的精确位置获取光谱信息。例如，在单个晶体水平上推出光学各向异性。样本的光谱和空间数据的生成允许对样品进行比仅光谱学或荧光显微镜更详细的调查。影响分辨率的大量可调参数起初可能看起来势不可挡。但是，制作检查表可能有助于熟悉系统。此技术需要手动调整映像硬件以及软件操作。必须直观地说明这两个方面，以展示这两种方法如何相辅相成。与纳尔逊·鲁塔霍加一起演示手术的将是艾米莉·罗德里格斯，我实验室的博士后研究员。要设置用于高光谱成像映射的成像器配置，从显微镜阶段开始，沿着发射光束路径向探测器移动，请将光学立方体的位置保留在光学显微镜旁边空位，然后将共聚焦显微镜光学立方体置于该位置，以引导样品通过可见光路径的发射。沿着光学路径向探测器移动，放置包含二色镜和滤波器的可见光学立方体，用于将可见发射定向到检测路径的位置。继续朝探测器的路径，将共通针孔光学立方体放在正确的位置，以引导光线穿过可见光检测路径并沿着此路径，将共分光谱仪光学立方体放在适当的位置，使发射的光到达探测器。当所有立方体都就位时，手动调整探测器的狭缝开口，以匹配所选针孔的大小。然后，在 PHySpec 软件中，选择针孔的孔径。对于欧洲双氟乙酰甲酸单晶的超光谱成像映射，手动将光学显微镜的20倍目标置于样品下，然后按下显微镜左侧的白色按钮以打开白光。转动按钮下方的旋钮以调整亮度。将显微镜右侧的前进旋钮设置为 R，将 20% 的信号发送到摄像机，将 80% 的信号发送到探测器。在 PHySpec 软件的彩色相机窗口中按播放以启动实时扫描。如果彩色相机窗口显示太暗或太黑的图像，请增加彩色相机选项卡下的曝光时间和/或增益值。如果图像太亮，请减少曝光时间和/或增益值。要聚焦样品，请转动显微镜的旋钮以调整舞台上目标之间的距离，并打开宽带灯快门以允许对样品进行紫外线激发。然后将强度旋钮转动到所需位置，以控制宽带灯激发的强度。单击"显示/隐藏比例图"可向晶体的亮场光学显微镜图像添加比例杆，并在紫外线全光或密闭照明下观察晶体图像。要在密闭照明或宽场照明之间进行选择，请使用斗杆和旋钮调整紫外灯场孔径的大小。在 SpectraPro SP 2300 选项卡下，选择一个波长来观察样品发射并调整探测器的曝光时间。要获取序列器中的高光谱立方体，请单击加号以添加新节点，然后单击"共和成像器"。单击多频谱采集并输入所需的 X 和 Y 位置计数以及所需的步进大小。选择相机同步和可见发射映射的硬件选项，然后单击"是否"。在音序器中，单击新添加的多频谱采集线以突出显示节点，然后单击"播放"以运行所选节点。要分析捕获的超光谱成像数据，例如，对于图像的光谱分布，请在"处理"菜单中选择"数据"和"裁剪"和"折弯"，以提高图像的信号与噪声比。对于发射强度配置文件，右键单击立方体图像并选择"创建 X 轮廓"以分析一条线。拖动以选择区域，然后右键单击感兴趣的区域。选择"添加到图形"。将发射强度显示为目标物理位置的函数。强度配置文件将显示在新图形中。要获取样本特定区域的发射频谱，请将光标悬停在立方体图像上，右键单击，然后单击矩形选择。拖动并单击以在所需区域上绘制选择形状。然后右键单击感兴趣的区域，然后选择"向图形添加选择"。在"添加到图形"窗口中，选择"创建新图形"以显示目标的排放光谱，然后单击"确定"。然后在选择新区域之前保存获取的频谱。此处显示了在正确焦点中调整样品后记录的晶体的亮场图像。可以清楚地观察到晶体的针状形态。在这里，可以观察到在紫外线激发下具有全照明或局部封闭照明的相同晶体的图像。受限照明可用于研究晶体中能量或光传递的影响，这些影响可能会触发波导一样的行为。例如，在此图像中，在非直接在激发下检测到强发射，这表明通过晶体进行有效的能量迁移。从获得的多光谱立方体中，也可以以图像的形式获得光谱分布，表示特定波长。特定发射波长的强度剖面和获得高光谱立方体任何像素或区域的发射光谱。例如，此分析的发射光谱显示了欧洲离子的最典型发射带。此外，沿不同晶体面的空间剖面表示尖端和侧面的发射更亮，可与三个空间方向的烷化物/烷化物离子距离相关。要在高光谱立方体的合理记录时间获得良好的信号，调整显微镜配置、对焦和探测器曝光时间非常重要。发射紫外线激发和可见发射，该技术可采用近红外激发和近红外发射检测。这允许它适用于大量的发光材料。光信号与一系列相关特征的相关性使得该技术对研究结构/属性关系非常有趣，包括纳米-生物相互作用的体外评估。

Research

JoVE Journal

Chemistry

Synthesis and Exfoliation of Discotic Zirconium Phosphates to Obtain Colloidal Liquid Crystals

合成与盘状磷酸盐锆剥落获取胶体液晶

Due to their abundance in natural clay and potential applications in advanced materials, discotic nanoparticles are of interest to scientists and ...

科研

化学

Coulomb Explosion Imaging as a Tool to Distinguish Between Stereoisomers

作为一种工具来区分立体异构体的库仑爆炸成像

This article shows how the COLTRIMS (Cold Target Recoil Ion Momentum Spectroscopy) or the "reaction microscope" technique can be used to distinguish ...

Measurements of Long-range Electronic Correlations During Femtosecond Diffraction Experiments Performed on Nanocrystals of Buckminsterfullerene

在纳米富勒的飞秒衍射实验的长程电子关联的测量

The precise details of the interaction of intense X-ray pulses with matter are a topic of intense interest to researchers attempting to interpret the ...

Electrochemical Impedance Spectroscopy as a Tool for Electrochemical Rate Constant Estimation

电化学阻抗谱作为电化学速率常数估计的一种工具

Electrochemical impedance spectroscopy (EIS) was used for advanced characterization of organic electroactive compounds along with cyclic voltammetry (CV). ...

Color Spot Test As a Presumptive Tool for the Rapid Detection of Synthetic Cathinones

色斑试验作为合成 Cathinones 快速检测的推定工具

Synthetic cathinones are a large class of new psychoactive substances (NPS) that are increasingly prevalent in drug seizures made by law enforcement and ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

表面光驱动分子电机的单分子成像

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

In situ FTIR Spectroscopy as a Tool for Investigation of Gas/Solid Interaction: Water-Enhanced CO2 Adsorption in UiO-66 Metal-Organic Framework

In situ FTIR Spectroscopy as a Tool for Investigation of Gas/Solid Interaction: Water-Enhanced CO2 Adsorption in UiO-66 Metal-Organic Framework

原位FTIR 光谱作为气体/固体相互作用研究工具：UiO-66 金属-有机框架中的水增强 CO2吸附

In situ infrared spectroscopy is an inexpensive, highly sensitive, and selective valuable tool to investigate the interaction of polycrystalline solids ...

Excitonic Hamiltonians for Calculating Optical Absorption Spectra and Optoelectronic Properties of Molecular Aggregates and Solids

用于计算分子聚合和固体光学吸收光谱和光电特性的外化学汉密尔顿

Rational design of disordered molecular aggregates and solids for optoelectronic applications relies on our ability to predict the properties of such ...

Mass Spectrometry-Guided Genome Mining as a Tool to Uncover Novel Natural Products

质谱导览基因组挖掘作为发现新天然产品的工具

The chemical space covered by natural products is immense and widely unrecognized. Therefore, convenient methodologies to perform wide-ranging evaluation ...

Atomic Force Microscopy Combined with Infrared Spectroscopy as a Tool to Probe Single Bacterium Chemistry

原子力显微镜与红外光谱相结合，作为探测单一细菌化学的工具

Atomic Force Microscopy-Infrared Spectroscopy (AFM-IR) is a novel combinatory technique, enabling simultaneous characterization of physical properties and ...