Quantitative Neutron Dark-Field Imaging of Milk: A Feasibility Study

Scattering studies of milk and milk products, which are highly relevant food products on the global market, are often utilized and reported in literature to investigate and understand the subtle microscopic structural differences between dairy samples. These structural features determine the physical properties and ultimately the texture of milk products and, thus, also influence the consumer’s experience. Small-angle neutron scattering is a prominent example, which enables observations of length scales, which convey proteins and fat globules in food-grade milk. In addition, deuteration enables contrast variations between the constituents of dairy products. In this study, we investigate the potential of probing small-angle neutron scattering from milk samples through quantitative neutron dark-field imaging using grating interferometry, to establish the feasibility of studying, in particular, fat globules and milk gel structures with this spatially resolved scattering technique.

Detection Of Surface Defects And Subsurface Defects Of Polished Optics With Multisensor Image Fusion

Surface defects (SDs) and subsurface defects (SSDs) are the key factors decreasing the laser damage threshold of optics. Due to the spatially stacked structure, accurately detecting and distinguishing them has become a major challenge. Herein a detection method for SDs and SSDs with multisensor image fusion is proposed. The optics is illuminated by a laser under dark field condition, and the defects are excited to generate scattering and fluorescence lights, which are received by two image sensors in a wide-field microscope. With the modified algorithms of image registration and feature-level fusion, different types of defects are identified and extracted from the scattering and fluorescence images. Experiments show that two imaging modes can be realized simultaneously by multisensor image fusion, and HF etching verifies that SDs and SSDs of polished optics can be accurately distinguished. This method provides a more targeted reference for the evaluation and control of the defects of optics, and exhibits potential in the application of material surface research.

Revealing the Quadrupole Radiation of Liquid Gallium Nanospheres

Gallium (Ga) nanospheres (NSs) with diameters ranging from 50 to 300 nm are fabricated by using femtosecond laser ablation. The forward scattering of large Ga nanospheres measured using dark-field microscopy is determined by the coherent interaction between dipole and quadrupole resonances while it becomes governed by the dipole resonance when evanescent wave excitation is employed. We demonstrate that the scattering spectrum and pattern of quadrupole of large Ga NS can be resolved by using a cross-polarized analyzer in the collection channel. The experimental observations agree well with the numerical simulation based on the complex refractive index of liquid Ga.

DNA mapping in the capsid of giant bacteriophage phiEL (Caudovirales: Myoviridae: Elvirus) by analytical electron microscopy

Introduction. Giant phiKZ-like bacteriophages have a unique protein formation inside the capsid, an inner body (IB) with supercoiled DNA molecule wrapped around it. Standard cryo-electron microscopy (cryo-EM) approaches do not allow to distinguish this structure from the surrounding nucleic acid of the phage. We previously developed an analytical approach to visualize protein-DNA complexes on Escherichia coli bacterial cell slices using the chemical element phosphorus as a marker. In the study presented, we adapted this technique for much smaller objects, namely the capsids of phiKZ-like bacteriophages.Material and methods. Following electron microscopy techniques were used in the study: analytical (AEM) (electron energy loss spectroscopy, EELS), and cryo-EM (images of samples subjected to low and high dose of electron irradiation were compared).Results. We studied DNA packaging inside the capsids of giant bacteriophages phiEL from the Myoviridae family that infect Pseudomonas aeruginosa. Phosphorus distribution maps were obtained, showing an asymmetrical arrangement of DNA inside the capsid.Discussion. We developed and applied an IB imaging technique using a high angle dark-field detector (HAADF) and the STEM-EELS analytical approach. Phosphorus mapping by EELS and cryo-electron microscopy revealed a protein formation as IB within the phage phiEL capsid. The size of IB was estimated using theoretical calculations.Conclusion. The developed technique can be applied to study the distribution of phosphorus in other DNA- or RNA-containing viruses at relatively low concentrations of the element sought.

Wnt11 family dependent morphogenesis during frog gastrulation is marked by the cleavage furrow protein anillin

Wnt11 family proteins are ligands that activate a type of Dishevelled-mediated, non-canonical Wnt signaling pathway. Loss of function causes defects in gastrulation and/or anterior-posterior axis extension in all vertebrates. Non-mammalian vertebrate genomes encode two Wnt11 family proteins whose distinct functions have been unclear. We knocked down zygotic Wnt11b and Wnt11, separately and together, in Xenopus laevis. Single morphants exhibited very similar phenotypes of delayed blastopore closure, but they had different phenotypes at the tailbud stage. In response to their very similar gastrulation phenotypes, we chose to characterize dual morphants. Using dark field illuminated time-lapse imaging and kymograph analysis, we identified a failure of dorsal blastopore lip maturation that correlated with slower blastopore closure and failure to internalize the endoderm at the dorsal blastopore lip. We connected these externally visible phenotypes to cellular events in the internal tissues – including the archenteron – by imaging intact embryos stained for anillin and microtubules. The cleavage furrow protein anillin provided an exceptional cytological marker for blastopore lip and archenteron morphogenesis and the consequent disruption through loss of Wnt11 signaling. These cytological changes suggest a novel role for the regulation of contractility and stiffness of the epithelial cells that result in dramatic shape changes and are important in gastrulation.

Advances in the Development of Phage-Based Probes for Detection of Bio-Species

Bacteriophages, abbreviated as “phages”, have been developed as emerging nanoprobes for the detection of a wide variety of biological species, such as biomarker molecules and pathogens. Nanosized phages can display a certain length of exogenous peptides of arbitrary sequence or single-chain variable fragments (scFv) of antibodies that specifically bind to the targets of interest, such as animal cells, bacteria, viruses, and protein molecules. Metal nanoparticles generally have unique plasmon resonance effects. Metal nanoparticles such as gold, silver, and magnetism are widely used in the field of visual detection. A phage can be assembled with metal nanoparticles to form an organic–inorganic hybrid probe due to its nanometer-scale size and excellent modifiability. Due to the unique plasmon resonance effect of this composite probe, this technology can be used to visually detect objects of interest under a dark-field microscope. In summary, this review summarizes the recent advances in the development of phage-based probes for ultra-sensitive detection of various bio-species, outlining the advantages and limitations of detection technology of phage-based assays, and highlighting the commonly used editing technologies of phage genomes such as homologous recombination and clustered regularly interspaced palindromic repeats/CRISPR-associated proteins system (CRISPR-Cas). Finally, we discuss the possible scenarios for clinical application of phage-probe-based detection methods.