Cryo-electron microscopy (cryo-EM) has for a long time been a technique of choice for determining structure of large and flexible macromolecular complexes that were difficult to study by other experimental techniques such as X-ray crystallography or nuclear magnetic resonance. However, a fast development of instruments and software for cryo-EM in the last decade has allowed that a large range o...

Human respiratory syncytial virus (hRSV) is an enveloped RNA virus that belongs to Paramyxoviridae family. HRSV is the most common cause of bronchiolitis and pneumonia in young children in the United States, and severe infections of infants and immuno-compromised adults can lead to death. In order to characterize the molecular mechanisms underlying hRSV assembly and viral and host cellular prot...

We present a modern, light portable device specifically designed for environmental samples for cryogenic transmission-electron microscopy (cryo-TEM) by on-site cryo-plunging. The power of cryo-TEM comes from preparation of artifact-free samples. However, in many studies, the samples must be collected at remote field locations, and the time involved in transporting samples back to the laboratory...

Here we present a protocol used to prepare cryo-TEM samples of Aspergillus niger spores, but which can easily be adapted for any number of microorganisms or solutions. We make use of a custom built cryo-transfer station and a modified cryo-SEM preparation chamber. The spores are taken from a culture, plunge-frozen in a liquid nitrogen slush and observed in the cryo-SEM to select a region of int...

The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assembl...

Cryo-electron tomography (cryo-ET) allows the visualization of supramolecular architecture in cells preserved in a close-to-physiological state. In order to supplement the structural information obtained by cryo-ET with the functional state of the molecules involved based on fluorescent labeling we developed a method of correlating light microscopy and cryo-ET. This method is suitable for inves...

Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.10.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous...

The aim of the study was to investigate distribution, enrichment and accumulation of heavy metals in soil and Trigonella foenum-graecum (var. Pusa Early Bunching) after fertigation with paper mill effluent. Doses of paper mill effluent viz. 5%, 10%, 25%, 50%, 75% and 100% were used for fertigation of T. foenum-graecum along with bore well water (control). The results revealed that paper mill ef...

The International Rice Research Institute (IRRI) Test Tube Mill and the Kett ”Pearlest” Polisher were evaluated in terms of milling performance in comparison with a McGill No. 2 Mill. Degree of milling (DOM), evaluated in terms of percent bran removal (the percentage of weight lost during milling) from brown rice, and whole kernel yield (WKY) were compared using sample lots of medium– and long–...

Healthy men produce an enormous number of sperms, far more than necessary for conception. However, several studies suggest that semen samples where the concentration of sperms is below 40 mill/mL may be associated with longer time to pregnancy or even subfertility, and specimens where the concentration of sperms is below 15 mill/mL may carry a high risk of infertility. Historic data from the 19...