Month: July 2016

Cross Sectioning of CuSn Connector of a Solar Cell

PurposeIon 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. GoalCross section to see the layer structure and their interfaces. Process Description(benchmark values for this particular sample)Mechanical pre-preparation with …

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Highly Selective Fluorescent and Colorimetric Probe for Live-cell Monitoring of Sulphide Based on Bioorthogonal Reaction

H2S is the third endogenously generated gaseous signaling compound and has also been known to involve a variety of physiological processes. To better understand its physiological and pathological functions, efficient methods for monitoring of H2S are desired. Azide fluorogenic probes are popular because they can take place bioorthogonal reactions. In this work, by employing a …

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Immuno – Electron Microscopy of High Pressure Frozen and Freeze Substituted Mouse Heart

Protocol HPF – AFS Mouse Heart IEM: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 (Leica Microsystems, Vienna, Austria). Freeze subs­titution was carried out using a Leica EM AFS (Leica Microsystems) in dry acetone containing …

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Cell Envelope Structure of a Gram-negative Thermotolerant y-Proteobacterium Acidithiobacillus sp., Strain HV2/2 and its Interaction with Pyrite

Sample Preparation: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 (Leica EM PACT2, Leica Microsystems GmbH, Vienna, Austria). Freeze substitution of the samples was performed in the automatic freeze subs­titution unit Leica EM AFS2 (Leica Microsystems GmbH, Vienna, Austria) according to standard …

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Translation Microscopy (TRAM) for Super-Resolution Imaging

Super-resolution microscopy is transforming our understanding of biology but accessibility is limited by its technical complexity, high costs and the requirement for bespoke sample preparation. We present a novel, simple and multi-color super-resolution microscopy technique, called translation microscopy (TRAM), in which a super-resolution image is restored from multiple diffraction-limited resolution observations using a conventional microscope …

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Symmetric Synapse – Clathrin Coated Endocytosis Pit in the Postsynaptic Dendrite

ProcedureWT 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 …

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Video Talk: CRISPR-Cas – From a Bacterial Adaptive Immune System to a Genome Engineering Tool

The CRISPR-Cas system was originally discovered as an adaptive immune system of bacteria and archaea to protect against viral attack. It is characterized by three major principles: Integration of foreign (viral) DNA into the bacterial CRISPR locus (Clustered Regularly Interspaced Short Palindromic Repeats) upon first infection Destruction of the labelled viral DNA by Cas (CRISPR …

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Electron Microscopy of High Pressure Frozen and Freeze Substituted Arabidopsis Thaliana Root Tips Cells

Protocol HPF – AFS for Morphological Analysis: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 (EM PACT; Leica Microsystems, Vienna, Austria). Freeze substitution was carried out using a Leica EM AFS2 (Leica Microsystems) in dry acetone containing 1% ddH2O, 1% OsO4 and 0.5% glutaraldehyde over …

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