The present study was aimed to assess the structural changes in protoscoleces of Echinococcus granulosus sensu stricto following exposure to different natural and chemical protoscolicidal agents using differential interference contrast (DIC)/Nomarski microscopy. Protoscoleces of sheep\'s liver cysts were collected aseptically. Individually, about 1000 protoscoleces were exposed to 0.5% silver nitrate, 20% hypertonic saline solution, 0.5% cetrimide solution and two different concentrations of garlic chloroformic extraction as well as phosphate-buffered saline (PBS). The protoscoleces viability was assessed using 0.1% eosin solution, and structural modifications in the protoscoleces were examined by DIC/Nomarski microscopy. The results revealed the degeneration of the tegument, disorganization of the hooks, and reduction of the size of the protoscoleces exposed to cetrimide, hypertonic sodium chloride, and silver nitrate. Furthermore, calcareous corpuscles became blurred and opaque and their numbers decreased in all the exposed samples except, those in PBS. The exposed protoscoleces to cetrimide and hypertonic sodium chloride solution showed extensive degeneration of the tegument and disorganization of the hooks. In the group exposed to 200 mg/ml chloroformic garlic extract, the protoscoleces\' width decreased. The length, width, and number of calcareous corpuscles also decreased significantly in the silver nitrate-exposed protoscoleces. The study concludes that protoscoleces exposed to different solutions; cetrimide 0.5% and hypertonic sodium chloride 20% caused more pronounced structural changes in the exposed protoscoleces. These changes were well demonstrated by DIC microscopy and can be used as a supplementary tool to evaluate the effects of protoscolicidal agents.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s12639-023-01632-4.

Lithium-carbon dioxide (Li-CO2 ) battery technology presents a promising opportunity for carbon capture and energy storage. Despite tremendous efforts in Li-CO2 batteries, the complex electrode/electrolyte/CO2 triple-phase interfacial processes remain poorly understood, in particular at the nanoscale. Here, using in situ atomic force microscopy and laser confocal microscopy-differential interference contrast microscopy, we directly observed the CO2 conversion processes in Li-CO2 batteries at the nanoscale, and further revealed a laser-tuned reaction pathway based on the real-time observations. During discharge, a bi-component composite, Li2 CO3 /C, deposits as micron-sized clusters through a 3D progressive growth model, followed by a 3D decomposition pathway during the subsequent recharge. When the cell operates under laser (λ=405 nm) irradiation, densely packed Li2 CO3 /C flakes deposit rapidly during discharge. Upon the recharge, they predominantly decompose at the interfaces of the flake and electrode, detaching themselves from the electrode and causing irreversible capacity degradation. In situ Raman shows that the laser promotes the formation of poorly soluble intermediates, Li2 C2 O4 , which in turn affects growth/decomposition pathways of Li2 CO3 /C and the cell performance. Our findings provide mechanistic insights into interfacial evolution in Li-CO2 batteries and the laser-tuned CO2 conversion reactions, which can inspire strategies of monitoring and controlling the multistep and multiphase interfacial reactions in advanced electrochemical devices.

Owing to the surge in plastic waste generated during the COVID-19 pandemic, concerns regarding microplastic pollution in aqueous environments are increasing. Since microplastics (MPs) are broken down into submicron (< 1 µm) and nanoscale plastics, their real-time morphological detection in water is necessary. However, the decrease in the scattering cross-section of MPs in aqueous media precludes morphological detection by bright-field microscopy. To address this problem, we propose and demonstrate a differential interference contrast (DIC) system that incorporates a magnification-enhancing system to detect MPs in aqueous samples. To detect MPs in both the stationary and mobile phases, a microfluidic chip was designed, taking into consideration the imaging depth of focus and flow resistance. MPs of various sizes flowing in deionized, tap, and pond water at varying speeds were observed under Static and Flow conditions. Successful real-time morphological detection and quantification of polystyrene beads down to 200 nm at a constant flow rate in water were achieved. Thus, the proposed novel method can significantly reduce analysis time and improve the size-detection limit. The proposed DIC microscopy system can be coupled with Raman or infrared spectroscopy in future studies for chemical composition analysis. ENVIRONMENTAL IMPLICATION: Microplastics (MPs), particularly submicron plastics < 1-µm, can pose a risk to human health and aquatic ecosystems. Existing methods for detecting MPs in the aqueous phase have several limitations, including the use of expensive instruments and prolonged and labor-intensive procedures. Our results clearly demonstrated that a new low-cost flow-channeled differential interference contrast microscopy enables the real-time morphological detection and quantification of MPs down to 200 nm under flowing conditions without sample labeling. Consequently, our proposed rapid method for accurate quantitative measurements can serve as a valuable reference for detecting submicron plastics in water samples.

Exfoliated flakes of layered materials, such as hexagonal boron nitride (hBN) and graphite with a thickness of several tens of nanometers, are used to construct van der Waals heterostructures. A flake with a desirable thickness, size, and shape is often selected from many exfoliated flakes placed randomly on a substrate using an optical microscope. This study examined the visualization of thick hBN and graphite flakes on SiO2/Si substrates through calculations and experiments. In particular, the study analyzed areas with different atomic layer thicknesses in a flake. For visualization, the SiO2thickness was optimized based on the calculation. As an experimental result, the area with different thicknesses in a hBN flake showed different brightness in the image obtained using an optical microscope with a narrow band-pass filter. The maximum contrast was 12% with respect to the difference of monolayer thickness. In addition, hBN and graphite flakes were observed by differential interference contrast (DIC) microscopy. In the observation, the area with different thicknesses exhibited different brightnesses and colors. Adjusting the DIC bias had a similar effect to selecting a wavelength using a narrow band-pass filter.

Various studies have been conducted to obtain quantitative phase information based on differential interference contrast (DIC) microscopy. As one such attempt, we propose in this study a single-shot quantitative phase imaging (QPI) method by combining two developments. First, an add-on optical system to a commercialized DIC microscope was developed to perform quantitative phase gradient imaging (QPGI) with single image acquisition using a polarization camera. Second, an algorithm was formulated to reconstitute QPI from the obtained QPGI by reducing linear artifacts, which arise in simply integrated QPGI images. To demonstrate the applicability of the developed system in cell biology, the system was used to measure various cell lines and compared with fluorescence microscopy images of the same field of view. Consistent with previous studies, nucleoli and lipid droplets can be imaged by the system with greater optical path lengths (OPL). The results also implied that combining fluorescence microscopy and the developed system might be more informative for cell biology research than using these methods individually. Exploiting the single-shot performance of the developed system, time-lapse imaging was also conducted to visualize the dynamics of intracellular granules in monocyte-/macrophage-like cells. Our proposed approach may accelerate the implementation of QPI in standard biomedical laboratories.

Collagen fibrils, linear arrangements of collagen monomers, 20-500 nm in diameter, comprising hundreds of molecules in their cross-section, are the fundamental structural unit in a variety of load-bearing tissues such as tendons, ligaments, skin, cornea, and bone. These fibrils often assemble into more complex structures, providing mechanical stability, strength, or toughness to the host tissue. Unfortunately, there is little information available on individual fibril dynamics, mechanics, growth, aggregation and remodeling because they are difficult to image using visible light as a probe. The principle quantity of interest is the fibril diameter, which is difficult to extract accurately, dynamically, in situ and non-destructively. An optical method, differential interference contrast (DIC) microscopy has been used to visualize dynamic structures that are as small as microtubules (25 nm diameter) and has been shown to be sensitive to the size of objects smaller than the wavelength of light. In this investigation, we take advantage of DIC microscopy\'s ability to report dimensions of nanometer scale objects to generate a curve that relates collagen diameter to DIC edge intensity shift (DIC-EIS). We further calibrate the curve using electron microscopy and demonstrate a linear correlation between fibril diameter and the DIC-EIS. Using a non-oil immersion, 40x objective (NA 0.6), collagen fibril diameters between ~100 nm to ~ 300 nm could be obtained with ±11 and ±4 nm accuracy for dehydrated and hydrated fibrils, respectively. This simple, nondestructive, label free method should advance our ability to directly examine fibril dynamics under experimental conditions that are physiologically relevant.

This paper describes how differences in the dynamics of targeting and nontargeting constructs can provide information on nanoparticle (NP)-cell interactions. We probed translational and rotational dynamics of functionalized Au nanostar (AuNS) nanoconstructs interacting with cells in serum-containing medium. We found that AuNS with targeting ligands had a larger dynamical footprint and faster rotational speed on cell membranes expressing human epidermal growth factor receptor 2 (HER-2) receptors compared to that of AuNS with nontargeting ligands. Targeting and nontargeting nanoconstructs displayed distinct membrane dynamics despite their similar protein adsorption profiles, which suggests that targeted interactions are preserved even in the presence of a protein corona. The high sensitivity of single-NP dynamics can be used to compare different nanoconstruct properties (such as NP size, shape, and surface chemistry) to improve their design as delivery vehicles.

Questions surrounding the optical properties of two-dimensional (2D) triangular single gold nanoplates (AuNPs) remain largely unanswered. Herein, a scanning-electron microscopy-correlated single-particle study was conducted to identify polarization-dependent optical properties of AuNPs under dark-field (DF) and differential interference contrast (DIC) microscopy. AuNPs with an aspect ratio of ∼3 showed a single broad DF scattering spectrum without separation of the two dipole and quadrupole resonance modes present in 2D AuNPs. Polarization-sensitive interference properties of the individual AuNPs were revealed through periodic changes in the intensities and types of DIC images obtained. A dipole resonance mode was found to mainly contribute to the polarization-sensitive interference properties of AuNPs. Furthermore, DIC polarization anisotropy allowed us to track the real-time orientation of a dipole resonance mode of a AuNP rotating on a live cell membrane.

Polyhydroxyalkanoates (PHAs), polyphosphate (poly-P) and polysulfide or elemental sulfur (poly-S) are the key functionally relevant polymers involved in the recently reported Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) process. However, little is known about the structural dynamics and storage states of these polymers. In particular, investigating the poly-S generated in this process is quite a superior challenge. This study was thus aimed at simultaneously qualitative-quantitative investigating poly-S and associated poly-P and PHAs through the integrated chemical analysis and Raman micro-spectroscopy coupled with multiple microscopic methods (i.e. optical microscopy, confocal laser scanning microscopy, and differential interference contrast microscopy). The chemical analytical results displayed a stable DS-EBPR phenotype in terms of sulfur conversion, P release/uptake and the dynamics of relevant polymers. The multiple microscopic images and Raman spectrum profiles further clearly demonstrated the existence of the polymers and their dynamic changes under alternating anaerobic-anoxic conditions, consistent with the chemical analytical results. In particular, Raman analysis for the first time unraveled the co-existence of S0/Sn2- species stored either intracellularly or extracellularly; and the dynamic conversions between S0/Sn2- and other sulfur species suggest that there might be a universal pool of bioavailable sulfur. The results reveal the mechanisms underlying the structural dynamics and changes in storage states of the relevant polymers that are functionally relevant to the carbon/phosphorus/sulfur-cycles during different metabolic phases. These mechanisms would otherwise not be obtained only using a traditional chemical analysis-based approach.

The cilium is a tiny organelle, with a length of 1-10 μm and a diameter of ~200 nm, that projects from the surface of many cells and functions to generate fluid flow and/or sense extracellular signals from the environment. Abnormalities in cilia may cause a broad spectrum of disease, i.e. the so-called ciliopathies. Multiple imaging approaches have been implemented to understand the structure, motion and function of the tiny cilium. In this review, we focus on the microscopic observations and analyses of the dynamic behaviors of both motile cilia and primary cilium. Motile cilia repeat reciprocal motions at 15-25 Hz with a clear asymmetry of effective and recovery strokes. Observing the fast movement of motile cilia requires a high-speed camera with a frame rate of more than 100 fps. The labeling of cilia tips enables the detailed analysis of the asymmetric beating motion of motile cilia. The primary cilium, which is imagined to be \'static,\' is also dynamic, i.e. it elongates, shrinks and disassembles, although this behavior is quite slower than that of motile cilia. The specific fluorescent labeling of primary cilium and time-lapse imaging are required to observe and analyze the slow behaviors of the primary cilium. We present some approaches, including some tips for successful procedures, in the successful imaging of the dynamic behaviors of motile cilia and primary cilium.