The effects of washing a collagen sample prior to TEM examination

Transmission electron microscopy (TEM) is an important analysis technique to visualize (bio)macromolecules and their assemblies, including collagen fibers. Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2-propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1 min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.     

Shape Factor Effect on Inclusion Sedimentation in Aluminum Melts

Metallurgical and Materials Transactions B - The quality requirements of aluminum products are steadily increasing, and the presence of non-metallic inclusions have a large impact on the quality of...     

Defining Dry Eye from a Clinical Perspective

Over the past decades, the number of patients with dry eye disease (DED) has increased dramatically. The incidence of DED is higher in Asia than in Europe and North America, suggesting the involvement of cultural or racial factors in DED etiology. Although many definitions of DED have been used, discrepancies exist between the various definitions of dry eye disease (DED) used across the globe. This article presents a clinical consensus on the definition of DED, as formulated in four meetings with global DED experts. The proposed new definition is as follows: “Dry eye is a multifactorial disease characterized by a persistently unstable and/or deficient tear film (TF) causing discomfort and/or visual impairment, accompanied by variable degrees of ocular surface epitheliopathy, inflammation and neurosensory abnormalities.” The key criteria for the diagnosis of DED are unstable TF, inflammation, ocular discomfort and visual impairment. This definition also recommends the assessment of ocular surface epitheliopathy and neurosensory abnormalities in each patient with suspected DED. It is easily applicable in clinical practice and should help practitioners diagnose DED consistently. This consensus definition of DED should also help to guide research and clinical trials that, to date, have been hampered by the lack of an established surrogate endpoint.     

Heat transport in dilute mixtures of3He in superfluid4He

We report new measurements of the effective thermal conductivity K_eff and relaxation time τ in dilute mixtures of³He in superfluid⁴He, with molar concentrationsX≤10⁻³. The temperature range extended fromT≈1.4 K toT _λ. Both K_cff and τ are found to agree with theoretical predictions, in contrast to previous experiments where significant differences were observed. A new thermal conductivity cell design was used which almost completely eliminates extraneous volumes and surfaces, and the earlier results are explained in relation to these design changes.     

A superconducting X-ray spectrometer with a tantalum absorber andlateral trapping

We report the development of Nb-Ta-Al-Al₂O₃-Al superconducting tunnel junction structures for high energy resolution and high efficiency X-ray detection. These devices utilize a Ta X-ray absorber with superconductor “bandgap engineered” quasiparticle trapping to improve charge collection. Experimental results at 0.3 K are presented, showing energy resolution of 102 eV full-width-half-maximum for 6 keV X-rays. Collected charge is in excess of 5×10⁶ electrons. The absorption efficiency is better than 35%. Devices thermally cycle with no change in characteristics