Hybrid pixel direct detector for electron energy loss spectroscopy.

We characterize a hybrid pixel direct detector and demonstrate its suitability for electron energy loss spectroscopy (EELS). The detector has a large dynamic range, narrow point spread function, detective quantum efficiency ≥ 0.8 even without single electron arrival discrimination, and it is resilient to radiation damage.

Design guidelines for an electron diffractometer for structural chemistry and structural biology.

3D electron diffraction has reached a stage where the structures of chemical compounds can be solved productively. Instrumentation is lagging behind this development, and to date dedicated electron diffractometers for data collection based on the rotation method do not exist. Current studies use transmission electron microscopes as a workaround.

Rapid Structure Determination of Microcrystalline Molecular Compounds Using Electron Diffraction.

Chemists of all fields currently publish about 50 000 crystal structures per year, the vast majority of which are X-ray structures. We determined two molecular structures by employing electron rather than X-ray diffraction. For this purpose, an EIGER hybrid pixel detector was fitted to a transmission electron microscope, yielding an electron diffractometer.

Cullin-RING ubiquitin E3 ligase regulation by the COP9 signalosome.

The cullin-RING ubiquitin E3 ligase (CRL) family comprises over 200 members in humans. The COP9 signalosome complex (CSN) regulates CRLs by removing their ubiquitin-like activator NEDD8. The CUL4A-RBX1-DDB1-DDB2 complex (CRL4A(DDB2)) monitors the genome for ultraviolet-light-induced DNA damage.

Oxidative doping renders graphene hydrophilic, facilitating its use as a support in biological TEM.

Graphene represents the first practical realization of crystalline supports in biological transmission electron microscopy (TEM) since their introduction over 30 years ago. The high transparency, minimal inelastic cross-section, and electrical conductivity of graphene are highly desirable characteristics for a TEM support. However, without a suitable method for rendering graphene supports, hydrophilic applications are limited.

Graphene: Substrate preparation and introduction.

This technical note describes the transfer of continuous, single-layer, pristine graphene to standard Quantifoil TEM grids. We compare the transmission properties of pristine graphene substrates to those of graphene oxide and thin amorphous carbon substrates. Positively stained DNA imaged across amorphous carbon is typically indiscernible and requires metal shadowing for sufficient contrast.

Preparation of 2D crystals of membrane proteins for high-resolution electron crystallography data collection.

Electron crystallography is a powerful technique for the structure determination of membrane proteins as well as soluble proteins. Sample preparation for 2D membrane protein crystals is a crucial step, as proteins have to be prepared for electron microscopy at close to native conditions. In this review, we discuss the factors of sample preparation that are key to elucidating the atomic structure of membrane proteins using electron crystallography.

Graphene oxide: a substrate for optimizing preparations of frozen-hydrated samples.

Graphene oxide is a hydrophilic derivative of graphene to which biological macromolecules readily attach, with properties superior to those of amorphous carbon films commonly used in electron microscopy. The single-layered crystalline lattice of carbon is highly electron transparent, and exhibits conductivity higher than amorphous carbon. Hence, graphene oxide is a particularly promising substrate for the examination of biological materials by electron microscopy.

Cryoelectron microscopy map of Atadenovirus reveals cross-genus structural differences from human adenovirus.

A three-dimensional (3D) cryoelectron microscopy reconstruction of the prototype Atadenovirus (OAdV [an ovine adenovirus isolate]) showing information at a 10.6-A resolution (0.5 Fourier shell correlation) was derived by single-particle analysis.

Bilateral edge filter: photometrically weighted, discontinuity based edge detection.

Edge-detection algorithms have the potential to play an increasingly important role both in single particle analysis (for the detection of randomly oriented particles), and in tomography (for the segmentation of 3D volumes). However, the majority of traditional linear filters are significantly affected by noise as well as artefacts, and offer limited selectivity. The Bilateral edge filter presented here is an adaptation of the Bilateral filter [Jiang, W.

Fast automated cell phenotype image classification.

Background: The genomic revolution has led to rapid growth in sequencing of genes and proteins, and attention is now turning to the function of the encoded proteins. In this respect, microscope imaging of a protein's sub-cellular localisation is proving invaluable, and recent advances in automated fluorescent microscopy allow protein localisations to be imaged in high throughput. Hence there is a need for large scale automated computational techniques to efficiently quantify, distinguish and classify sub-cellular images.