Name: obtaining 3d chemical maps by energy filtered transmission electron microscopy tomography
Uploaded: 2018-06-09T12:30:00
Description: Watch this Scientific Journal Video about Obtaining 3D Chemical Maps by Energy Filtered Transmission Electron Microscopy Tomography at JoVE.com

We are grateful to the French Ministry of Higher Education and Research, Conventions Industrielles de Formation par la Recherche (CIFRE) and IFP Energies Nouvelles for their financial support.

1. Sample Preparation
<ol>
	<li>Crush the sample in a mortar and disperse it in alcohol or distilled water; place a droplet of the sample on a microscopy grid and let it dry. 
	NOTE: Samples such as silica alumina or titania alumina can be a powder or an extruded material, and can be crushed and dispersed in a solution using ultrasound. In general, for ET analysis, it is important that the sample concentration on the grid is low, to avoid sample superposition and shadowing when tilting the grid at large angles. Th...

<ol>
	<li>Frank, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electron Tomography - Methods for Three-Dimensional Visualization of Structures in the Cell. , (2006).</li><li>Midgley, P. A., Dunin-Borkowski, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+tomography+and+holography+in+materials+science.">Electron tomography and holography in materials science.</a> Nat. Mater. 8, 271-280 (2009).</li><li>Carenco, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+core+contribution+of+transmission+electron+microscopy+to+functional+nanomaterials+engineering.">The core contribution of transmission electron microscopy to functional nanomaterials engineering.</a> Nanoscale. 8 (3), 1260-1279 (2016).</li><li>Radon, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uber+die+Bestimmung+von+Funktionen+durch+ihre+Integralwerte+langs+gewisser+Mannigfaltigkeiten.">Uber die Bestimmung von Funktionen durch ihre Integralwerte langs gewisser Mannigfaltigkeiten.</a> Akad. Wiss. 69, 262-277 (1917).</li><li>Radermacher, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radon+transform+techniques+for+alignment+and+three-dimensional+reconstruction+from+random+projections.">Radon transform techniques for alignment and three-dimensional reconstruction from random projections.</a> Scanning Microscopy. 11, 171-177 (1997).</li><li>Roiban, L., Sorbier, L., Pichon, C., Pham-Huu, C., Drillon, M., Ersen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D-TEM+investigation+of+the+nanostructure+of+a+&#948;-Al2O3+catalyst+support+decorated+with+Pd+nanoparticles.">3D-TEM investigation of the nanostructure of a &#948;-Al2O3 catalyst support decorated with Pd nanoparticles.</a> Nanoscale. 4 (3), 946-954 (2012).</li><li>Georgescu, D., Roiban, L., Ersen, O., Ihiawakrim, D., Baia, L., Simon, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+on+Ag+doped+porous+TiO2+nanostructures:+a+comprehensive+study+of+their+structural+and+morphological+characteristics.">Insights on Ag doped porous TiO2 nanostructures: a comprehensive study of their structural and morphological characteristics.</a> RSC Adv. 2 (12), 5358 (2012).</li><li>Shakeri, M., Roiban, L., Yazerski, V., Prieto, G., Gebbink, M. J. M. G., de Jongh, P. E., de Jong, K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+and+Sizing+Nanoreactors+To+Confine+Metal+Complexes+for+Enhanced+Catalytic+Performance.">Engineering and Sizing Nanoreactors To Confine Metal Complexes for Enhanced Catalytic Performance.</a> ACS Catal. 4 (10), 3791-3796 (2014).</li><li>Midgley, P. A., Weyland, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+electron+microscopy+in+the+physical+sciences:+the+development+of+Z-contrast+and+EFTEM+tomography.">3D electron microscopy in the physical sciences: the development of Z-contrast and EFTEM tomography.</a> Ultramicroscopy. 96 (3-4), 413-431 (2003).</li><li>Ersen, O., Florea, I., Hirlimann, C., Pham-Huu, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+nanomaterials+with+3D+electron+microscopy.">Exploring nanomaterials with 3D electron microscopy.</a> Mater. Today. 18 (7), 395-408 (2015).</li><li>Lepinay, K., Lorut, F., Pantel, R., Epicier, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+3D+tomography+of+28nm+high+K+metal+gate+transistor:+STEM+XEDS+experimental+method+and+results.">Chemical 3D tomography of 28nm high K metal gate transistor: STEM XEDS experimental method and results.</a> Micron. 47, 43-49 (2013).</li><li>Roiban, L., Sorbier, L., Pichon, C., Bayle-Guillemaud, P., Werckmann, J., Drillon, M., Ersen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Chemistry+of+Multiphase+Nanomaterials+by+Energy-Filtered+Transmission+Electron+Microscopy+Tomography.">Three-Dimensional Chemistry of Multiphase Nanomaterials by Energy-Filtered Transmission Electron Microscopy Tomography.</a> Microsc. Microanal. 18 (05), 1118-1128 (2012).</li><li>Roiban, L., Sorbier, L., Hirlimann, C., Ersen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3+D+Chemical+Distribution+of+Titania-Alumina+Catalyst+Supports+Prepared+by+the+Swing-pH+Method.">3 D Chemical Distribution of Titania-Alumina Catalyst Supports Prepared by the Swing-pH Method.</a> ChemCatChem. 8 (9), 1651-1657 (2016).</li><li>Egerton, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electron Energy-Loss Spectroscopy in the Electron Microscope. , (2011).</li><li>Messaoudi, C., Aschman, N., Cunha, M., Oikawa, T., Sorzano, C. O. S., Marco, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Chemical+Mapping+by+EFTEM-TomoJ+Including+Improvement+of+SNR+by+PCA+and+ART+Reconstruction+of+Volume+by+Noise+Suppression.">Three-Dimensional Chemical Mapping by EFTEM-TomoJ Including Improvement of SNR by PCA and ART Reconstruction of Volume by Noise Suppression.</a> Microscopy and Microanalysis. 19 (6), 1669-1677 (2013).</li><li>Pettersen, E. F., Goddard, T. D., Huang, C. C., Couch, G. S., Greenblatt, D. M., Meng, E. C., Ferrin, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=UCSF+Chimera-A+visualization+system+for+exploratory+research+and+analysis.">UCSF Chimera-A visualization system for exploratory research and analysis.</a> Journal of Computational Chemistry. 25 (13), (2004).</li><li>Roiban, L., Ersen, O., Hirlimann, C., Drillon, M., Chaumonnot, A., Lemaitre, L., Gay, A. S., Sorbier, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Analytical+Surface+Quantification+of+Heterogeneous+Silica-Alumina+Catalyst+Supports.">Three-Dimensional Analytical Surface Quantification of Heterogeneous Silica-Alumina Catalyst Supports.</a> ChemCatChem. 9 (18), 3503-3512 (2017).</li><li>Kremer, J. R., Mastronarde, D. N., McIntosh, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computer+visualization+of+three-dimensional+image+data+using+IMOD.">Computer visualization of three-dimensional image data using IMOD.</a> J Struct Biol. 116 (1), 71-76 (1996).</li><li>Align RGB planes. ImageJ Available from: <a target="_blank" href="https://ImageJ.net/Align_RGB_planes">https://ImageJ.net/Align_RGB_planes</a> (2018)</li><li>MessaoudiI, C., Boudier, T., Sorzano, C., Marco, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TomoJ:+tomography+software+for+three-dimensional+reconstruction+in+transmission+electron+microscopy.">TomoJ: tomography software for three-dimensional reconstruction in transmission electron microscopy.</a> BMC Bioinf. 8 (1), 288 (2007).</li><li>Saxton, W. O., Baumeister, W., Hahn, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+reconstruction+of+imperfect+two-dimensional+crystals.">Three-dimensional reconstruction of imperfect two-dimensional crystals.</a> Ultramicroscopy. 13 (1-2), 57-70 (1984).</li></ol>

The aim of this paper is to describe how to obtain 3D chemical maps using EFTEM tomography. This protocol is completely original and was developed by the authors.
EFTEM tomography as described here has several drawbacks: (i) Only samples that are electron beam resistant can be analyzed, due to the long exposure time needed for obtaining filtered images. (ii) EFTEM tomography is sensitive to the diffraction contrast. (iii) Many of the alignments were performed manually. To obtain the 3D chemica...

The properties of functional materials are dependent on their 3D parameters. To fully comprehend their properties and enhance their functions, it is important to analyze their morphology and chemical distribution in 3D. Electron tomography1 (ET) is one of the best techniques to provide this information at the nanometer scale2,3. It consists of rotating the sample over a large angular range and recording one image at each angular step. The obtained tilt series is used to reconstruct the volume of the sample by using mathematical algorithms based on the Radon transform4,5. Selecting grey levels in the volume helps to model the sample in 3D and quantify 3D parameters like particle localization6 and size distribution7, pore position and size distribution8, etc.
In general, ET is performed with an electron microscope by tilting the sample to the maximum possible angle, preferably more than 70&#176; in either direction. At each tilt angle, a projection of the sample is recorded forming an images tilt series. That tilt series is aligned and used to reconstruct the volume of the sample which will be segmented and quantified. Because the sample cannot be rotated from -90&#176; to +90&#176;, the reconstructed volume has an anisotropic resolution along the orthogonal axis9 due to the blind recording angle.
ET can be performed in different imaging modes. The bright field TEM mode (BF-TEM) is used to study amorphous materials, biological samples, polymers, or catalyst supports with complex shapes. The image analysis is based on the differentiation of the gray levels characterizing the density of the components10 (a dense component will be more dark than a lighter, i.e., less dense component). High-angle annular dark field in scanning TEM mode (HAADF-STEM) is used to analyze crystalline samples. The signal provides chemical information as a function of the atomic number; a heavy component of the sample will appear brighter that a lighter one9. Other modes, like Energy Dispersive X-ray spectroscopy (EDX), which collects the X-ray emitted by the material11, and energy filtered imaging mode (EFTEM)12,13, are also capable of assessing the 3D chemical distribution within the sample.
In EFTEM imaging, the 2D chemical maps can be recorded using a TEM with an electron energy spectrometer. The spectrometer acts as a magnetic prism by dispersing the electrons as a function of their energy. An image is created by the electrons depending on the energy lost from interacting with a specific atom. If the same 2D chemical map is computed at different tilt angles, a tilt series of chemical projections is obtained, which can be used to reconstruct the 3D chemical volume.
Not all the materials can be analyzed by EFTEM tomography. The technique is reserved for samples with weak or disordered materials. Nevertheless, it can be used for analyzing light elements that are very difficult to differentiate when using other imaging techniques. In addition, to obtain reliable 2D chemical maps, the thickness of the material is required to be less than the mean free path of the electrons through the material14. Under this condition, the probability of having a single electron interacting with a single atom is greatest. Two methods are used to calculate a 2D chemical map. The first one, and the most used is the &#34;three-windows method&#34;, where two filtered energy windows are recorded before the ionization edge of the element under analysis, and a third after the ionization edge13. The first two images are used to estimate the background, which is extrapolated using a power law at the position of the third window and subtracted from it. The obtained image is the projection of the 3D distribution of the analyzed chemical element in the sample volume. The second method is called the &#34;jump-ratio&#34;; it uses only two energy-filtered images, one before and one after the ionization edge. This method is qualitative, as the final image is calculated only by performing the ratio between those two images, and does not account for background energy variation.
By combining EFTEM with ET, the analytical tomography of the filtered energy can be obtained. EFTEM tomography and atom probe tomography (APT) are complementary techniques. As compared to APT, EFTEM tomography is a non-destructive characterization analysis that does not need complex sample preparation. It can be used to perform various characterizations on a unique nanoparticle. EFTEM tomography can analyze insulating materials, while APT needs at the very least laser assistance to measure them. APT runs at the atomic scale, while EFTEM tomography performs adequately with a lower resolution. EFTEM tomography is pertinent only for samples that resist beam degradation during the experiment. To record all the filtered images at all the tilted angles, the sample can be exposed to the electron beam for as long as 2 h. Moreover, to record a maximum chemical signal in the 2D maps, longer exposition durations at high beam intensity may be necessary. In such conditions, the beam sensitive samples suffer drastic morphological and chemical changes. Therefore, a precise measurement of the sample sensitivity to the electron beam must be established before the experiment. In addition, EFTEM tomography is the result of recording as many tomograms as necessary to determine the spatial location and nature of the chemical elements that are present in the sample. Nevertheless, EFTEM tomography can provide important information concerning the 3D chemical distribution for samples, such as catalyst supports, to give new insights for modeling their catalytic applications.
Today it is possible to use dedicated software that can select the energy interval, record filtered energy window images, and calculate the chemical maps at different tilt angles. They allow tilting the sample, tracking, focusing, and recording the filtered image in EFTEM mode. The 2D chemical maps can be calculated, and then the tilt series can be aligned, the chemical volume computed using iterative algorithms, and finally the series can be segmented and quantified15,16.

<table>
	<tbody>
		<tr>
			<td>JEOL 2100f</td>
			<td>JEOL</td>
			<td></td>
			<td>Electron microscope</td>
		</tr>
		<tr>
			<td>Tridiem Gatan Imaging Filter (GIF)</td>
			<td>Gatan</td>
			<td></td>
			<td>Post colum energy filter</td>
		</tr>
		<tr>
			<td>Digital micrograph</td>
			<td>Gatan</td>
			<td></td>
			<td>Software</td>
		</tr>
		<tr>
			<td>Gatan EFTEM tomography plugin</td>
			<td>Gatan</td>
			<td></td>
			<td>Dedicated software to record filtered tilt series for EFTEM tomograohy</td>
		</tr>
		<tr>
			<td>Tomoj</td>
			<td>Imagej plugin http://www.cmib.fr/en/download/softwares/</td>
			<td></td>
			<td>Free software developed by Currie Institute in Paris, France for electron tomography</td>
		</tr>
		<tr>
			<td>EFTEM-Tomoj</td>
			<td>Imagej plugin http://www.cmib.fr/en/download/softwares/</td>
			<td></td>
			<td>Free software developed by Currie Institute in Paris, France , for EFTEM imaging</td>
		</tr>
		<tr>
			<td>Imod</td>
			<td>http://bio3d.colorado.edu/imod/</td>
			<td></td>
			<td>Free software developed by University of Colorado, USA for electron tomography</td>
		</tr>
		<tr>
			<td>Imagej</td>
			<td>https://imagej.nih.gov/ij/</td>
			<td></td>
			<td>Free software developed by National Institute of Mental Health, Bethesda, Maryland, USA for images treatment</td>
		</tr>
		<tr>
			<td>Merge channels</td>
			<td>https://imagej.net/Color_Image_Processing</td>
			<td></td>
			<td>Fonction in Imagej allowing to give different colors to volumes while they are overlapped</td>
		</tr>
		<tr>
			<td>3D Slicer</td>
			<td>https://www.slicer.org/</td>
			<td></td>
			<td>Free software developed by a large consortium lead by Ron Kikinis , Harvard Medical School, Boston, MA, SUA</td>
		</tr>
		<tr>
			<td>Chimera</td>
			<td>https://www.cgl.ucsf.edu/chimera/</td>
			<td></td>
			<td>Free software developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco,for data segmentation, cuatification and visualisation of 3D models</td>
		</tr>
		<tr>
			<td>silica alumina support of catalyst</td>
			<td>IFPEN</td>
			<td></td>
			<td>sample prepared for eleboration of this protocol</td>
		</tr>
		<tr>
			<td>titania alumina support of catalyst</td>
			<td>IFPEN</td>
			<td></td>
			<td>sample prepared for eleboration of this protocol</td>
		</tr>
		<tr>
			<td>alcohol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>water</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Au nanoparticles of 5 nm</td>
			<td>BBI Solutions</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Holey carbn film 200 mesh microscopy grid</td>
			<td>Agar</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EDX sepctrometer</td>
			<td>Oxford Instruments</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

obtaining 3d chemical maps by energy filtered transmission electron microscopy tomography

Energy filtered transmission electron microscopy tomography (EFTEM tomography) can provide three-dimensional (3D) chemical maps of materials at a nanometric scale. EFTEM tomography can separate chemical elements that are very difficult to distinguish using other imaging techniques. The experimental protocol described here shows how to create 3D chemical maps to understand the chemical distribution and morphology of a material. Sample preparation steps for data segmentation are presented. This protocol permits the 3D distribution analysis of chemical elements in a nanometric sample. However, it should be noted that currently, the 3D chemical maps can only be generated for samples that are not beam sensitive, since the recording of filtered images requires long exposure times to an intense electron beam. The protocol was applied to quantify the chemical distribution of the components of two different heterogeneous catalyst supports. In the first study, the chemical distribution of aluminum and titanium in titania-alumina supports was analyzed. The samples were prepared using the swing-pH method. In the second, the chemical distribution of aluminum and silicon in silica-alumina supports that were prepared using the sol-powder and mechanical mixture methods was examined.

An example of the application of this protocol is shown in reference13. EFTEM tomography was used for analyzing titania alumina catalyst supports. To enhance the catalytic activity of the active phase of MoS2 nanoparticles, in applications like hydrodesulfurization (HDS), it is important that titania is preponderant at the support surface, and in contact with the active phase. It is known that titania has a smaller specific surface than alumina. The aim ...

Watch this Scientific Journal Video about Obtaining 3D Chemical Maps by Energy Filtered Transmission Electron Microscopy Tomography at JoVE.com

Obtaining 3D Chemical Maps by Energy Filtered Transmission Electron Microscopy Tomography

This paper describes a protocol to achieve 3D chemical maps combining energy filtered imaging and electron tomography. The chemical distribution of two catalyst supports formed by elements that are difficult to distinguish by other imaging techniques was studied. Each application consists of mapping overlapped chemical elements - respectively spaced-ionization edges.

Energy filtered transmission electron microscopy tomography (EFTEM tomography) can provide three-dimensional (3D) chemical maps of materials at a ...

The overall goal of this protocol is to obtain, visualize and quantify the 3D chemical maps obtained by energy filtered electron tomography. This method can help answer key questions in material science, chemistry and catalysis fields, such as how the components of a given material are mixed at nanometric scale. The main advantage of this technique is that it provides information concerning the chemical distribution in the bulk and at the surface of the sample.Energy filtered imaging can separate chemical elements that are very difficult to distinguish using other imaging techniques. The experimental protocol described here shows the means of obtaining 3D chemical maps but also it tries to provide ideas to quantify some of the 3D chemical parameters. The 3D chemical maps can be only generated for samples that are not beam sensitive since the recording of filtered images requires long exposure times to relatively intense electron beam.To begin prepare a sample such as silica alumina or titania alumina, as described in the accompanying text protocol. Next use an electron microscope to record energy filtered tilt series. These images will be used to calculate each chemical map of the search chemical element.Open the three filered tilt series consisting of two pre-edge series and one post-edge series in image J for analysis. Now open the specialized EFTEMTomoJ plugin and click on then click on Image or Stack to select the tilt series. In the table that appears enter the energy at which the tilt series was recorded as the energy shift.Also enter the slit width and the exposure time of each filtered image. Check all three tilt series under mapping and ensure that they will be used for computing the chemical maps using the power law. Then click on Next.Choose the first image as the reference image and then click Apply followed by Next. At this point a preview image of the proposed alignment appears. Visually verify that the three images recorded at the same tilt angle are well superimposed, showing no shift between the images.This protocol was performed on the version 0.9 of the EFTEMTomoJ plugin. At this moment the filtered images recorded at the same tilt angle are aligned. Next select the EFTEMTomoJ window, the images corresponding to the background and to the chemical edge.Then select the model of the signal extraction as power and click on Create Map. Save the tilt series of the chemical map and then repeat this process for all of the other chemical tilt series. In order to quantify this drift, first open the zero loss aligned tilt series.Then open the chemical maps aligned tilt series, go to Image, Color and select Merge Channels. Select the file corresponding to zero loss for the red color, the first chemical element for green. Uncheck Create Composite and check Keep Source Image.At this point a stack is created at each tilt angle for all the recorded images. Next go to Plugins and select Align RGB Plans. Here red is the reference image.Select green and using the arrows overlap it over the red one. Once in position click on Next and repeat this process for all of the other angles. Go to Image, down to Color and select Split Channels to split the RGB stack into tow stacks.Red corresponds to zero loss and green corresponds to the chemical maps with the drift corrected. Finally be sure to save the tilt series, then repeat the actions for the second chemical element and remember to save the file. For 3D chemical mapping again sit at a computer and open image J.Open the TomoJ plugin by going to plugins and selecting TomoJ.This will select the load angle form file. Because all the tilt series are already aligned navigate directly to reconstruction and calculate the volumes of zero loss as well as the chemical volumes using a reconstruction algorithm. It is recommended to use an interactive algorithm to reconstruct the chemical volumes.Also using this software it is possible to reconstruct the volumes using the GPU of your computer. Once all volumes are computed use the Merge Channels options to apply different colors to the obtained volumes and overlap them in a single volume to obtain the 3D chemical map. Finally follow along in the accompanying text protocol for additional details on 3D modeling and quantification of the chemical information, surface area and pore size distribution.EFTEM Tomography was used here to analyze three different samples of titania alumina catalyst supports with different component ratios. It was found that titania at high concentrations forms clusters that are embedded in the alumina. These models show the chemical distribution of titania and alumina on the surface of the samples.It was found that independent of the proportion of titania and alumina in the sample, the surface of the sample is covered with titania in a proportion of about 30%Alumina has a high specificity to the surface and the goal is to have titania supported by alumina. After modeling it was found that the sample containing 10%titania has a layer that is only about 10 nanometers thick on the surface. This variance could play a role in the sample's catalytic applications.It is important to mention that energy filtered electron tomography is a time consuming technique. While attempting this procedure remember that energy filtered imaging is sensitive to the defraction contrast, but also the thickness of the sample and its resistance to the electron beam must be verified. After its development this technique paved the way for researchers in materials science to better correlate the morphological and chemical parameters with the properties of that studied material.We hope that after following this protocol the new users of electron tomography will have a better understanding of the step to follow and on the difficulties encountered in reconstruct a volume, segment a model and quantify its parameters.

Research

JoVE Journal

Chemistry

Quantitative Detection of Trace Explosive Vapors by Programmed Temperature Desorption Gas Chromatography-Electron Capture Detector

The direct liquid deposition of solution standards onto sorbent-filled thermal desorption tubes is used for the quantitative analysis of trace explosive ...

1,3,5-Triphenylbenzene and Corannulene as Electron Receptors for Lithium Solvated Electron Solutions

The authors report on conductivity studies carried out on lithium solvated electron solutions (LiSES) prepared using two types of polyaromatic ...

A Simple, Low-cost, and Robust System to Measure the Volume of Hydrogen Evolved by Chemical Reactions with Aqueous Solutions

There is a growing research interest in the development of portable systems which can deliver hydrogen on-demand to proton exchange membrane (PEM) ...

Facile Preparation of (2Z,4E)-Dienamides by the Olefination of Electron-deficient Alkenes with Allyl Acetate

Facile Preparation of 2Z,4E-Dienamides by the Olefination of Electron-deficient Alkenes with Allyl Acetate

Direct cross-coupling between two alkenes via vinylic C-H bond activation represents an efficient strategy for the synthesis of butadienes with high ...

Quantifying X-Ray Fluorescence Data Using MAPS

The quantification of X-ray fluorescence (XRF) microscopy maps by fitting the raw spectra to a known standard is crucial for evaluating chemical ...

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 ...

Analysis of Minerals Produced by hFOB 1.19 and Saos-2 Cells Using Transmission Electron Microscopy with Energy Dispersive X-ray Microanalysis

This video presents the use of transmission electron microscopy with energy dispersive X-ray microanalysis (TEM-EDX) to compare the state of minerals in ...

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

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 ...

Preparation of Graphene-Supported Microwell Liquid Cells for In Situ Transmission Electron Microscopy

Preparation of Graphene-Supported Microwell Liquid Cells for In Situ Transmission Electron Microscopy

The fabrication and preparation of graphene-supported microwell liquid cells (GSMLCs) for in situ electron microscopy is presented in a stepwise protocol. ...

Photodeposition of Pd onto Colloidal Au Nanorods by Surface Plasmon Excitation

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