Leica Science Lab - Tag : Application Note https://www.leica-microsystems.com//science-lab/tag/tags/application-note/show/Tag/ Article tagged with Application Note en-US https://www.leica-microsystems.com/20352 EM Sample Preparation Macroscale to Nanoscale Pore Analysis of Shale and Carbonate Rocks Physical porosity in rocks, like shale and carbonate, has a large effect on the their storage capacity. The pore geometries also affect their permeability. Imaging the visible pore space provides insights into the physical pore space, pore geometries, and the associated mineral and organic matter phases relevant for storage and transport. For example, organic-matter porosity is an important property of mature organic-rich shales providing storage capacity for liquids and gases, like hydrocarbons. Imaging the microstructure is the only technique which delivers data giving direct insight into the organic matter porosity. However, imaging porosity in fine-grained shale as well as carbonate rocks in a representative way at high resolution is challenging. https://www.leica-microsystems.com//science-lab/macroscale-to-nanoscale-pore-analysis-of-shale-and-carbonate-rocks/ Tue, 04 Sep 2018 22:00:00 +0000 PhD Jop Klaver, PhD Joyce Schmatz, MSc Mingze Jiang, PhD James DeRose https://www.leica-microsystems.com/18094 EM Sample Preparation Micro-CT of Insect Larva Protocol Species: red blood worm (midge larva) Critical point drying of midge larvae with subsequent X-ray micro-computed tomography (micro-CT) to reconstruct the inner anatomy. https://www.leica-microsystems.com//science-lab/micro-ct-of-insect-larva-protocol/ Tue, 14 Mar 2017 07:14:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/18221 EM Sample Preparation Contrast Enhancement of Polycrystalline Metals - Sample Preparation for SEM Application Note - Ion milling is a perfect alternative for chemical etching, especially for polycrystalline metals, such as copper. Ion milling can be used to increase the contrast of the grain structure and their interfaces. In contrast to chemical etching the milling process is clean, safe and easy to operate. Ion energy and milling time depend on the milling rate of the metal. https://www.leica-microsystems.com//science-lab/contrast-enhancement-of-polycrystalline-metals-sample-preparation-for-sem/ Wed, 01 Feb 2017 10:33:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/19176 EM Sample Preparation Drosophila larvae - Sample Preparation for Cryo-SEM Application Note for Leica EM ACE900 - Drosophila larvae were sandwiched between two 3 mm aluminum slit carriers with the 100 μm cavities facing each other and high-pressure frozen with a Leica EM HPM100. No ethanol as synchronization media was used, 1-hexadecene was used as filler. The wholes of the slit carriers were filled with filter tips dipped in 1-hexadecene to keep the carrier sandwich complete after freezing. https://www.leica-microsystems.com//science-lab/drosophila-larvae-sample-preparation-for-cryo-sem/ Mon, 30 Jan 2017 09:50:00 +0000 Dr. Andres Kaech, Prof. Damian Brunner https://www.leica-microsystems.com/19179 EM Sample Preparation Giardia lamblia - Sample Preparation for Cryo-SEM Application Note for Leica EM ACE900 - A 100 mesh copper grid (12 um thickness) was dipped into a concentrated Giardia suspension and sandwiched between two flat 3 mm aluminum specimen carriers with scratched surfaces. Subsequently, the sandwich was transferred to the widened hole of a middle plate (3.1 mm diameter). A 50 um spacer ring was added on top and the specimen immediately frozen with an HPM100 high-pressure freezing machine without using alcohol as synchronization fluid. https://www.leica-microsystems.com//science-lab/giardia-lamblia-sample-preparation-for-cryo-sem/ Mon, 30 Jan 2017 09:35:00 +0000 Dr. Andres Kaech, Joe Paulin Zumthor https://www.leica-microsystems.com/18156 EM Sample Preparation Cross Sectioning of Cadmiumsulphide (CdS) for Cathodoluminescence Cathodoluminescence can be used to achieve spectra and high resolution images of impurity and structural defects in semicondoctors, minerals and insulating materials. This application note explains how to prepare a perfect sample surface for carhodoluminescence and how to use ion beam slope cutting to prepare the sample surface free of any preparation artefacts. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-cadmiumsulphide-cds-for-cathodoluminescence/ Tue, 24 Jan 2017 18:12:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18207 EM Sample Preparation Picea abies (L.) KARST - Sample Preparation for TEM Application Note for Leica EM AMW - Plants (5-years old) were grown in pots filled with soil and kept in greenhouse conditions. Five weeks before harvesting the plants were transferred into growth chambers and cultivated at a temperature of 20°C during daytime and 12°C overnight. The relative humidity was set at 60% and the photoactive radiation was 500 μmol m-2 s-1 during daytime. Sample preparation for transmission electron microscopy (TEM) was performed in order to develop a standard protocol that would reduce sample preparation time for TEM-investigations. Therefore the overall and fine structure of leaf cells prepared with the Leica EM AMW were compared with leaf cells that were prepared with a conventional fixation protocol at room temperature. https://www.leica-microsystems.com//science-lab/picea-abies-l-karst-sample-preparation-for-tem/ Mon, 23 Jan 2017 19:24:00 +0000 Prof. Bernd Zechmann, Prof. Günther Zellnig https://www.leica-microsystems.com/18196 EM Sample Preparation Cross Sectioning of Copper for Electron Backscattered Diffraction (EBSD) Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating diffraction patterns (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphological studies. The information depth is just a few nm, therefore good sample preparation is very important to avoid damages. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-copper-for-electron-backscattered-diffraction-ebsd/ Thu, 19 Jan 2017 16:37:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18247 EM Sample Preparation Porous Ceramics - Sample Preparation for SEM Application Note for Leica EM RES102 - Ceramic membrane filters with pore sizes down to a few nanometres must be investigated in cross-section with regard to the structure of the pores. The smallest pores are of special interest. In most cases, conventional grinding methods cannot be used for such problems, as the pore structure would be distorted. This applies in particular to the pores in the nanometre range. https://www.leica-microsystems.com//science-lab/porous-ceramics-sample-preparation-for-sem/ Tue, 17 Jan 2017 07:32:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18319 EM Sample Preparation Maple (Acer saccharum) Leaves - High Pressure Freezing and Freeze Substitution for TEM Application Note for Leica EM HPM100 - Leaves were immersed in hexadecene and placed under a gentle (0.3 bar) vacuum for 10 minutes to evacuate the internal air spaces. The leaves were then trimmed to fit the carriers and placed in the 200 μm side of a 6 mm Type A specimen carrier. Free space was filled with additional hexadecene after which a 6 mm Type B specimen carrier was placed on top with the flat side down. https://www.leica-microsystems.com//science-lab/maple-acer-saccharum-leaves-high-pressure-freezing-and-freeze-substitution-for-tem/ Mon, 16 Jan 2017 17:08:00 +0000 Dr. Kim Rensing https://www.leica-microsystems.com/18249 EM Sample Preparation Removal of Surface Layers - Sample Preparation for SEM and TEM Application Note for Leica EM RES102 - Sometimes it is necessary to remove surface layers to gain access to the real surface structure. That can be a native oxide, or layers coming from the preparation process itself, like re-deposition. Depending on the layers thickness and the energy used for the cleaning process, it takes between a few seconds and half an hour. The energy depends on the milling rate of the material. https://www.leica-microsystems.com//science-lab/removal-of-surface-layers-sample-preparation-for-sem-and-tem/ Wed, 11 Jan 2017 10:10:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18354 EM Sample Preparation Cross Section of Solar Cells Application Note for Leica EM TIC020, Leica EM TIC 3X - Cross section of a complete solar cell. https://www.leica-microsystems.com//science-lab/cross-section-of-solar-cells/ Fri, 23 Dec 2016 06:47:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18362 EM Sample Preparation Cross Sectioning of a Multilayer System - Preparation of a Perfect Sample Surface for EBSD Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating a diffraction pattern (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphology studies. The information depth is just a few nm. Therefore good sample preparation is very important to avoid any damage. This is very difficult in case of multilayer system with big differences in hardness. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-a-multilayer-system-preparation-of-a-perfect-sample-surface-for-ebsd/ Thu, 22 Dec 2016 20:48:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18317 EM Sample Preparation High-Pressure Freezing and Freeze Substitution of Hep-2 Cells Infected with Chlamydia pneumoniae Application Note for Leica EM HPM100 - Hep-2 cells infected with Chlamydia pneumoniae were cultured on carbon-coated 6 mm Sapphire discs. Cells were high-pressure frozen in an EM HPM100 using the 6 mm CLEM middle plate with following setup: Sapphire disc with cells, spacer 200 μm, bare Sapphire disc, 2 spacers 200 μm. Ethanol was used as a synchronization fluid to transfer pressure at room temperature prior to cooling. https://www.leica-microsystems.com//science-lab/high-pressure-freezing-and-freeze-substitution-of-hep-2-cells-infected-with-chlamydia-pneumoniae/ Tue, 20 Dec 2016 11:03:00 +0000 Dr. Andres Kaech https://www.leica-microsystems.com/18243 EM Sample Preparation Multilayer Systems with Widely Different Sputter Rates - Sample Preparation for TEM Application Note for Leica EM RES102 - The multi-layer system to be prepared in cross-section consists of a Si substrate, a TiN layer with a thickness of a few nm and a 500 nm W layer. All these components have extreme differences in their hardness, their atomic weight and in their sputter rates. A preparation of this kind of samples with sample rotation would lead to a wall overlying the area of the layers. https://www.leica-microsystems.com//science-lab/multilayer-systems-with-widely-different-sputter-rates-sample-preparation-for-tem/ Fri, 16 Dec 2016 16:57:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18225 EM Sample Preparation In-Containing Compound Semiconductors - Sample Preparation for TEM Application Note for Leica EM RES102 - Previous studies showed that surface accumulation of In occurs when InP was milled in a conventional way with Ar ions. The consequence is In islands on the sample surface. This leads to low quality of TEM samples. To remove these islands, reactive ion milling with iodine ions (RIBE / CAIBE) can be used. This method has the disadvantage of polluting the ion guns and the vacuum system of the ion milling device and leads to chemical reactions with the sample material. To avoid these problems we prepared these samples very gently with low energy Ar ions. https://www.leica-microsystems.com//science-lab/in-containing-compound-semiconductors-sample-preparation-for-tem/ Mon, 12 Dec 2016 10:13:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/19003 EM Sample Preparation Carbon Coating for Polymeric Materials Application Note fo Leica EM ACE600 - A solid understanding of polymer property-structure relationships is critical to improve and shorten development routes to new products. A direct way to determine correlations between structure and mechanical properties is provided by electron microscopy. Electron microscopy techniques have an important advantage over other methods, as they can provide local information at high spatial resolution. However, a major problem with polymers is their inherent lack of contrast. https://www.leica-microsystems.com//science-lab/carbon-coating-for-polymeric-materials/ Mon, 05 Dec 2016 09:22:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18312 EM Sample Preparation Commercially Available Hand Creams - Sample Preparation for Cryo-SEM Application Note for Leica EM HPM100 - Hand creams having different water contents were applied into the 100 μm cavities of two 3 mm type A sample carriers which were then closed cream sides inwards. The sample assembly was high pressure frozen with a Leica EM HPM100 and moved to a cooled Leica EM VCT100 loading station. https://www.leica-microsystems.com//science-lab/commercially-available-hand-creams-sample-preparation-for-cryo-sem/ Fri, 18 Nov 2016 17:22:00 +0000 Dr. Kim Rensing https://www.leica-microsystems.com/18255 EM Sample Preparation "Shallow Trench Isolation" Structures - Sample Preparation for TEM Application Note for Leica EM RES102 - The cross-sectional preparation of structured semiconductor materials requires a very thorough mechanical pre-preparation. In doing this, it must be ensured that the structure of interest should be located as close to the centre of the sample as possible. As the sample will be ion milled from both sides, a specific preparation of the structure is necessary in most cases, which means that you must thin these structures from both sides. https://www.leica-microsystems.com//science-lab/shallow-trench-isolation-structures-sample-preparation-for-tem/ Fri, 18 Nov 2016 16:50:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18110 EM Sample Preparation Tobacco Leaf - Critical Point Drying Protocol for SEM Application Note for Leica EM CPD300 - Critical point drying of tobacco leafs with subsequent platinum coating and SEM analysis. https://www.leica-microsystems.com//science-lab/tobacco-leaf-critical-point-drying-protocol-for-sem/ Fri, 18 Nov 2016 16:21:00 +0000 Dr. Martin W. Goldberg, M.Sc. Christine Richardson https://www.leica-microsystems.com/18245 EM Sample Preparation Paper Samples - Sample Preparation for SEM Application Note for Leica EM RES102 - A coated paper sample has been prepared with ion beam slope cutting in order to test the procedure with regard to its applicability. With the use of ion beam slope cutting a cross section of paper could be prepared. On the basis of this sample processing, it was possible to show the largely unaffected original structure of the thermally-sensitive paper in the scanning electron microscope. https://www.leica-microsystems.com//science-lab/paper-samples-sample-preparation-for-sem/ Wed, 16 Nov 2016 14:01:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18241 EM Sample Preparation Metal Films and Sheets - Sample Preparation for TEM Application Note for Leica EM RES102 - Most metal films already have a thickness that requires no further mechanical pre-preparation. Frequently, however, they are also domed, which can lead to undefined milling angles. This is a disadvantage, particularly for films that contain inclusions, and that therefore mostly require very flat milling angles. Metal sheets are thicker than 100 µm. Mechanical pre-preparation is necessary to obtain an acceptable initial thickness and a good surface quality for ion milling. https://www.leica-microsystems.com//science-lab/metal-films-and-sheets-sample-preparation-for-tem/ Wed, 16 Nov 2016 13:32:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18106 EM Sample Preparation Nematode E. dianae - Critical Point Drying Protocol for SEM Application Note for Leica EM CPD300 - Critical point drying of nematode Eubostrichus dianae to detect the ectosymbiotic bacteria layer with subsequent gold coating ans SEM analysis. https://www.leica-microsystems.com//science-lab/nematode-e-dianae-critical-point-drying-protocol-for-sem/ Wed, 16 Nov 2016 11:05:00 +0000 Mag. Nikolaus Leisch https://www.leica-microsystems.com/18940 EM Sample Preparation Visualization of Membrane Dynamics with Millisecond Temporal Resolution Application Note for Leica EM ICE, Leica EM AFS2 - Electrical stimulation of neurons combined with high-pressure freezing allows physiological activation of synaptic activity and precise control over the time frame of the induced synaptic activity. https://www.leica-microsystems.com//science-lab/visualization-of-membrane-dynamics-with-millisecond-temporal-resolution/ Mon, 07 Nov 2016 10:53:00 +0000 PhD Shigeki Watanabe https://www.leica-microsystems.com/18213 EM Sample Preparation Cross-Sectional Preparation of Structured Semiconductor Materials for TEM Application Note for Leica EM RES102 - The vertical layer construction of a semiconductor structure should be examined as a TEM cross-sectional sample. In addition to the specific preparation of the desired structure, the widely different sputter rates and atomic weights of the individual components represent the level of difficulty involved with this preparation problem. https://www.leica-microsystems.com//science-lab/cross-sectional-preparation-of-structured-semiconductor-materials-for-tem/ Mon, 31 Oct 2016 08:15:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18235 EM Sample Preparation Contrast Enhancement of Polished Cross Sections of Semiconductor Structures - Sample Preparation for SEM Application Note for Leica EM RES102 - The surfaces of polished cross sections often show fine scratches and residues of the removed material or of the abrasive material. The artefacts are strongly material-dependent, and are mostly only detectable at higher resolutions in the scanning electron microscope. A further problem arises from the fact that the ground section mostly only has low contrast, i.e., in the structures of the semiconductor materials are very difficult to discern. With the use of ion beam milling, the ground sections of semiconductor structures can be "contrasted". https://www.leica-microsystems.com//science-lab/contrast-enhancement-of-polished-cross-sections-of-semiconductor-structures-sample-preparation-for-sem/ Fri, 28 Oct 2016 12:08:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18142 EM Sample Preparation Wall Cress Pod Protocol - Critical Point Drying of Arabidopsis thaliana for SEM Application Note for Leica EM CPD300 - Critical point drying of wall cress (Arabidopsis thaliana) pod with subsequent gold coating and SEM analysis. https://www.leica-microsystems.com//science-lab/wall-cress-pod-protocol-critical-point-drying-of-arabidopsis-thaliana-for-sem/ Tue, 25 Oct 2016 08:00:00 +0000 Dr. Chen LiYu https://www.leica-microsystems.com/18257 EM Sample Preparation Surface Modification of ZnAg Sample - Sample Preparation for SEM Application Note for Leica EM RES102 - By means of cleaning, polishing and contrast enhancement a soft ZnAg sample should be prepared to obtain information concerning the grain structure and interfaces of the sample. The sample is contaminated after mechanical polishing. There are still some scratches on the surface. Grain structure is almost invisible. https://www.leica-microsystems.com//science-lab/surface-modification-of-znag-sample-sample-preparation-for-sem/ Mon, 17 Oct 2016 08:16:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18364 EM Sample Preparation Cross Sectioning of Ni/Cu on Steel for EBSD Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating a diffraction pattern (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphology studies. The information depth is just a few nm. Therefore a good sample preparation is very important. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-nicu-on-steel-for-ebsd/ Fri, 14 Oct 2016 09:45:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18096 EM Sample Preparation Bacteria Protocol - Critical Point Drying of E. coli for SEM Application Note for Leica EM CPD300 - Critical point drying of E. coli with subsequent platinum / palladium coating and SEM analysis. Sample was inserted into a filter disc (Pore size: 16 - 40 μm) and placed into the filter discs and porous pots holder. Cultivate fungi and bacteria on agar containing growth medium for 3 days. https://www.leica-microsystems.com//science-lab/bacteria-protocol-critical-point-drying-of-e-coli-for-sem/ Thu, 13 Oct 2016 10:04:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/18178 EM Sample Preparation Cross Sectioning of Painted Concrete Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve flat cuts of material combinations consisting of hard and soft materials. Here, cross sectioning of painted concrete is describes in order to visualize the interface between paint and concrete. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-painted-concrete/ Mon, 10 Oct 2016 20:39:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18104 EM Sample Preparation Clawed Frog Nuclear Envelope Protocol Application Note for Leica EM CPD300 - Critical point drying of nuclear pores from clawed frog oocytes with subsequent chromium coating and SEM analysis. Silicon chips containing the samples were placed into the filter discs and porous pots holder. https://www.leica-microsystems.com//science-lab/clawed-frog-nuclear-envelope-protocol/ Thu, 06 Oct 2016 16:00:00 +0000 Dr. Martin W. Goldberg, M.Sc. Christine Richardson https://www.leica-microsystems.com/18092 EM Sample Preparation Micro-Computed Tomography (micro-CT) of Insect Brain Protocol Application Note for Leica EM CPD300 - Critical point drying of the blow fly with subsequent X-ray micro-computed tomography (micro-CT) to detect neuroanatomical features. https://www.leica-microsystems.com//science-lab/micro-computed-tomography-micro-ct-of-insect-brain-protocol/ Thu, 06 Oct 2016 14:41:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/18259 EM Sample Preparation Thin Metal Foils with Coatings - Sample Preparation for SEM Application Note for Leica EM RES102 - Thin foils are mostly unstable because of their thickness of a few microns. This makes it difficult to do slope cutting without any protection of the sample. A common realisation to protect the sample surface is by sticking a cover glass on top of the sample. Another issue is cutting the foils before ion milling. The sample edge should be flat and sharp without any broken areas. A razor blade is mostly the best solution. A protected sample can salso be sawed with a wire saw. https://www.leica-microsystems.com//science-lab/thin-metal-foils-with-coatings-sample-preparation-for-sem/ Wed, 05 Oct 2016 07:16:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18818 EM Sample Preparation Ultra-thin Carbon Support Films for Improved STEM-EELS Analysis of Nanoparticles Application Note for Leica EM ACE600 - Recent developments in aberration corrected transmission electron microscopes as well as further improvements in monochromaters and spectrometers have pushed the attainable energy resolution for Electron energy loss spectroscopy (EELS) to 100 meV and beyond. STEM-EELS of individual nanomaterials can be challenging due the necessity of a support film. https://www.leica-microsystems.com//science-lab/ultra-thin-carbon-support-films-for-improved-stem-eels-analysis-of-nanoparticles/ Wed, 21 Sep 2016 11:27:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18820 EM Sample Preparation Ways to Reveal More from your Samples: Ultra-Thin Carbon Films Application Note for Leica EM ACE600 - Much of the battle involved in obtaining good transmission electron microscopy data is in the specimen preparation itself. Even though some nanomaterials are already electron transparent (e.g. nanoparticles and proteins) and often do not require further thinning procedures, they need to be dispersed onto thin support films. https://www.leica-microsystems.com//science-lab/ways-to-reveal-more-from-your-samples-ultra-thin-carbon-films/ Wed, 21 Sep 2016 11:23:00 +0000 PhD Frédéric Leroux, Jan de Weert https://www.leica-microsystems.com/18192 EM Sample Preparation Cross Sectioning of a Superconductive Wire Application Note for Leica EM TIC 3X - Purpose: The shape of the wire is difficult for ion beam slope cutting. Goal: Cross sectional preparation to see the structure of the wire. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-a-superconductive-wire/ Tue, 20 Sep 2016 16:41:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18149 EM Sample Preparation Structural Study of C. elegans Application Note for Leica EM ICE, Leica EM AFS2 - Wildtype L4 stage C. elegans (N2 strain) were placed in the 100 μm deep side of Lecithin-coated (see detailed protocol*) type A 3 mm Cu/Au carriers (Leica) with extracellular filler containing 1% (w/v) Agarose type IX and 2% (w/v) Bovine Serum Albumin in bacteria medium (see preparation details**) and sandwiched with the flat side of Lecithin-coated type B 3 mm Cu/Au carriers (Leica). Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/structural-study-of-c-elegans/ Fri, 16 Sep 2016 08:15:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. Rob Mesman https://www.leica-microsystems.com/18344 EM Sample Preparation Cross Section of an Aluminium Sample for Electron Backscattered Diffraction (EBSD) Application Note for Leica EM TIC020, Leica EM TIC 3X - Electron backscattered diffraction (EBSD) is for example used to examine the crystallographic orientation of material. The sample preparation for such samples is sometimes very tricky as the depth of information is just few nm (~20nm or less). That means the sample surface must be flat and free of preparation artefacts. Mechanical polishing leads mostly to sample surfaces damages. https://www.leica-microsystems.com//science-lab/cross-section-of-an-aluminium-sample-for-electron-backscattered-diffraction-ebsd/ Thu, 15 Sep 2016 07:54:00 +0000 Robert Ranner https://www.leica-microsystems.com/18301 EM Sample Preparation Freeze-Fracture Replication of Pyramidal Cells Application Note for Leica EM HPM100 - Frozen samples (90 μm thick slices frozen by HPM100) were inserted into a double replica table and then fractured into two pieces at –130°C (after insertion of the tissue into BAF 060 the samples should be left in the chamber for 20 min to reach the –130°C). https://www.leica-microsystems.com//science-lab/freeze-fracture-replication-of-pyramidal-cells/ Thu, 08 Sep 2016 16:23:00 +0000 Akos Kulik https://www.leica-microsystems.com/18239 EM Sample Preparation Ion Beam Polishing of Sample Surfaces - Sample Preparation for SEM Application Note for Leica EM RES102 - Ion milling can be used to reduce the roughness of sample surfaces. Small angles less than 6° with respect to the sample surface are necessary. The high voltage depends on the material to be prepared. The reason for the levelling effect is the different milling angle of flat and rough surface areas. The milling rate is lower for small angles. The rough surface area will be faster milled. Ion polishing is often the final step of sample preparation. The prerequisite is a perfect mechanical prepreparation as samples with deep surface scratches cannot be ion polished. Soft materials usually have a smeared sample surface after mechanical polishing. It is necessary to remove this smeared material before ion polishing. Otherwise the above mentioned polishing effect does not work. https://www.leica-microsystems.com//science-lab/ion-beam-polishing-of-sample-surfaces-sample-preparation-for-sem/ Tue, 06 Sep 2016 11:11:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18215 EM Sample Preparation Cleaning of Smeared Sample Surfaces - Sample Preparation for SEM Application Note for Leica EM RES102 - Mechanical polishing of soft materials or hard / soft material combinations is tricky. The mechanical polishing process leads very often to smearing of the soft material. The smeared material covers the surface and fills small pores or holes. Grain structures, interfaces and other structural details can be masked. An additional ion milling step with milling angles between 10° C and 15° C with respect to the sample surface can remove or reduce the contamination. https://www.leica-microsystems.com//science-lab/cleaning-of-smeared-sample-surfaces-sample-preparation-for-sem/ Fri, 02 Sep 2016 06:49:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18090 EM Sample Preparation Micro-Computed Tomography (micro-CT) of Book Scorpion Musculature Application Note for Leica EM CPD300 - Critical point drying of book scorpion with subsequent X-ray micro-computed tomography (micro-CT) to detect anatomical features with special regard to the musculature. https://www.leica-microsystems.com//science-lab/micro-computed-tomography-micro-ct-of-book-scorpion-musculature/ Thu, 01 Sep 2016 08:39:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/18146 EM Sample Preparation Cultured Rat Hippocampal Neurons Application Note for Leica EM ICE - Rat Hippocampal neurons, cultured on 50 μm thick Aclar (Aclar embedding film, EMS) for 19 days, were frozen in the 100 μm deep side of lecithin coated (detailed protocol Appendix I) type A 3 mm Cu/Au carriers (Leica) and sandwiched with the flat side of lecithin coated type B 3 mm Cu/Au carriers (Leica). No additional filler was used, only cell culture medium with the addition of Hepes buffer pH 7.2 to a final concentration of 25 mM. Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/cultured-rat-hippocampal-neurons/ Mon, 29 Aug 2016 07:12:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. W. H. Mueller, Prof. Dr. C. C. Hoogenraad https://www.leica-microsystems.com/18000 EM Sample Preparation Epoxy Resin Embedding of Animal and Human Tissues for Pathological Diagnosis and Research Application Note for Leica EM AMW - The tissues were fixed in the modified Karnovsky fixative generally by immersion overnight (at minimum 4h at room temperature). Then pieces of approx. 1mm3 were cut with a sharp razor blade and processed for embedding in the Leica EM AMW Microwave Tissue Processor. https://www.leica-microsystems.com//science-lab/epoxy-resin-embedding-of-animal-and-human-tissues-for-pathological-diagnosis-and-research/ Fri, 19 Aug 2016 08:34:00 +0000 Dr. Josef A. Schroeder https://www.leica-microsystems.com/18209 EM Sample Preparation Ceramics - Sample Preparation for TEM Application Note for Leica EM RES102 - Ceramic samples are mostly very brittle, and are therefore very difficult to thin mechanically to a low starting thickness for ion beam milling. The ion beam milling of insulators often leads to static charging of the surface of the sample. This, in turn, reduces the sputter rate. When using the Ti standard holder (standard TEM holder), however, sufficient secondary electrons are created by the ion beam also falling on the sample holder to largely compensate for the static charging. https://www.leica-microsystems.com//science-lab/ceramics-sample-preparation-for-tem/ Mon, 15 Aug 2016 18:46:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18295 EM Sample Preparation Cryo-Electron Microscopy of Vitreous Sections (CEMOVIS) of Yeast Application Note for Leica EM HPM100 - The sections are of yeast frozen with a Leica HPM100 high pressure freezer in the copper tube system, the cell paste was mixed with a pH 6.5 MES/dextran buffer so that a final MES concentration of 50mM and a dextran concentration of 20% was achieved. https://www.leica-microsystems.com//science-lab/cryo-electron-microscopy-of-vitreous-sections-cemovis-of-yeast/ Fri, 12 Aug 2016 14:51:00 +0000 Dr. Jonathan O'Driscoll, Dr. Daniel Kofi Clare, Prof. Helen Saibil https://www.leica-microsystems.com/18144 EM Sample Preparation Wrinkled Giant Hyssop Leaf Protocol Application Note for Leica EM CPD300 - Critical point drying of wrinkled giant hyssop leaf with subsequent gold coating and SEM analysis. https://www.leica-microsystems.com//science-lab/wrinkled-giant-hyssop-leaf-protocol/ Thu, 11 Aug 2016 17:04:00 +0000 Dr. Guo JianSheng https://www.leica-microsystems.com/18100 EM Sample Preparation Black Mould Protocol Application Note for Leica EM CPD300 - Critical point drying of Black mould (Aspergilus niger) with subsequent platinum / palladium coating and SEM analysis to detect conidiospores. Sample was inserted into a filter disc (Pore size: 16 - 40 μm) and placed into the filter discs and porous pots holder. https://www.leica-microsystems.com//science-lab/black-mould-protocol/ Tue, 09 Aug 2016 12:01:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/18151 EM Sample Preparation Cross Sectioning of Alumina Application Note for Leica EM TIC 3X - Alumina is very difficult to handle and almost impossible to prepare with conventional methods. Ion beam slope cutting is a method that can achieve cross sections of material combinations consisting of hard and soft components. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-alumina/ Mon, 08 Aug 2016 07:48:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/17998 EM Sample Preparation Arabidopsis thaliana(L.) Accession Col Application Note for Leica EM AMW - Plants were grown in growth chambers under defined conditions. After stratification for 4 days at 4°C seeds were grown in pots with soil with 9/15 hours day/night photoperiod. Day and night temperatures were 22°C and 18°C, respectively, the relative humidity was 60% and the plants were kept at 100% relative soil water content. Light intensity varied between 110 and 140 μmol m-2 s-1. https://www.leica-microsystems.com//science-lab/arabidopsis-thalianal-accession-col/ Mon, 01 Aug 2016 13:03:00 +0000 Prof. Bernd Zechmann, Prof. Günther Zellnig https://www.leica-microsystems.com/18190 EM Sample Preparation Cross Sectioning of CuSn Connector of a Solar Cell Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve flat cross sections of soft materials or material combinations consisting of hard and soft components. The CuSn connector is very soft. Mechanical polishing leads to smearing. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-cusn-connector-of-a-solar-cell/ Fri, 29 Jul 2016 11:56:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/17987 EM Sample Preparation Immuno - Electron Microscopy of High Pressure Frozen and Freeze Substituted Mouse Heart Application Note for Leica EM AFS2 - Mouse heart tissue from wild-type (WT) and αT -catenin KO (KO) mice was excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer Leica EM PACT. Freeze subs­titution was carried out using a Leica EM AFS in dry acetone containing 2% ddH2O and 0.1% glutaraldehyde over a 4-days period. https://www.leica-microsystems.com//science-lab/immuno-electron-microscopy-of-high-pressure-frozen-and-freeze-substituted-mouse-heart/ Wed, 27 Jul 2016 15:35:00 +0000 Riet de Rycke https://www.leica-microsystems.com/17979 EM Sample Preparation Cell Envelope Structure of a Gram-negative Thermotolerant y-Proteobacterium Acidithiobacillus sp., Strain HV2/2 and its Interaction with Pyrite Application Note for Leica EM AFS2 - Cells of the the moderately thermophilic Acidithiobacillus sp.strain HV2/2, were centrifuged at 20,0009xg and then cryo-immobilized by high-pressure freezing on gold carriers. https://www.leica-microsystems.com//science-lab/cell-envelope-structure-of-a-gram-negative-thermotolerant-y-proteobacterium-acidithiobacillus-sp-strain-hv22-and-its-interaction-with-pyrite/ Mon, 25 Jul 2016 10:12:00 +0000 Dr. Andreas Klingl https://www.leica-microsystems.com/18049 EM Sample Preparation Neuroscience Symmetric Synapse - Clathrin Coated Endocytosis Pit in the Postsynaptic Dendrite Application Note for Leica EM ICE - WT hippocampal neurons were plated at a density of 80,000 cells/cm2 on 6 mm sapphire disks for 14 days. Sample were frozen using a high-pressure freezer (Leica EM ICE) under a pressure of 2100bar by mounting it into a sandwich support with extracellular solution containing 15% of Ficoll 400, to assess ice crystal damage. The Cryo-fixation was achieved within milliseconds allowing simultaneous immobilization of all macromolecular components. After freezing, sam­ple was transferred into cryovials containing 1% glutaraldehyde, 1% osmium tetroxide, 1% milliQwater in anhydrous acetone and processed in an automated freeze-substitution device (Leica EM AFS2). https://www.leica-microsystems.com//science-lab/symmetric-synapse-clathrin-coated-endocytosis-pit-in-the-postsynaptic-dendrite/ Wed, 20 Jul 2016 08:56:00 +0000 Dr. Shuwen Chang https://www.leica-microsystems.com/17975 EM Sample Preparation Electron Microscopy of High Pressure Frozen and Freeze Substituted Arabidopsis Thaliana Root Tips Cells Application Note for Leica EM AFS2 - Arabidopsis thaliana roots (mutant PIN1pro:PIN1-GFP;bex5-1) were excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer. Freeze substitution was carried out using a Leica EM AFS2. https://www.leica-microsystems.com//science-lab/electron-microscopy-of-high-pressure-frozen-and-freeze-substituted-arabidopsis-thaliana-root-tips-cells/ Fri, 08 Jul 2016 10:02:00 +0000 Riet de Rycke https://www.leica-microsystems.com/18368 EM Sample Preparation Cross Sectioning of Oil Shale Rock Application Note for Leica TIC 3X - High quality sample preparation of large area to investigate the sample in the SEM. For the mechanical preparation step diamond lapping foils of 9μm subsequently 2μm and finally 0.5μm grain size were used. It took about 1.5 hours. The sample was removed from the stub with a razor blade after TXP processing and fixed onto the holder of the rotary stage of the Leica EM TIC 3X. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-oil-shale-rock/ Tue, 28 Jun 2016 13:04:00 +0000 Robert Ranner https://www.leica-microsystems.com/18284 EM Sample Preparation Cryo-SEM Imaging of Latex Paint Application Note for Leica EM VCT100, Leica EM ACE600 - A thin layer of latex paint was spread on a clean, scored, silicon chip. The sample was immediately plunge frozen in liquid ethane and transferred under LN2 to the Leica EM VCT100 loading station where it was placed in the customized sample holder. https://www.leica-microsystems.com//science-lab/cryo-sem-imaging-of-latex-paint/ Thu, 16 Jun 2016 16:50:00 +0000 Dr. Levi Felts, Dr. Kim Rensing, Chris Frethem https://www.leica-microsystems.com/18184 EM Sample Preparation Cross Sectioning of Rubber (Tire) Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve cross sections of soft materials or material combinations consisting of hard and soft components. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-rubber-tire/ Tue, 14 Jun 2016 10:30:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18086 EM Sample Preparation Water Flea Protocol Application Note for Leica EM CPD300 - Critical point drying of water flea with subsequent gold coating and SEM-Analysis to detect finde surface structures. https://www.leica-microsystems.com//science-lab/water-flea-protocol/ Tue, 07 Jun 2016 07:06:00 +0000 Mag. Dr. Daniela Gruber https://www.leica-microsystems.com/18079 EM Sample Preparation Human Blood Cells Protocol Application Note for Leica EM CPD300 - Life Science Research. Species: Human (Homo sapiens) Critical point drying of human blood with subsequent platinum / palladium coating and SEM analysis. https://www.leica-microsystems.com//science-lab/human-blood-cells-protocol/ Tue, 24 May 2016 08:03:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/17971 EM Sample Preparation Cross Sectioning of Basalt Fibres Application Note for Leica EM TIC 3X - Material Research. Purpose: The fibres are embedded in a soft matrix. That makes it difficult to prepare a cross section. Goal: Cross section of the basalt fibres. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-basalt-fibres/ Fri, 20 May 2016 15:31:00 +0000 PhD Wolfgang Grünewald