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Super sulphated cement (SSC) is a very promising substitute for traditional construction materials (i.e. Portland cement), due to its enhanced durability and particularly low environmental impact. This paper explores the microstructure and certain properties of SSC, focusing on the particular complexities of its microstructure and the difficulties of microanalysis of its hydrates. To do so, SSC paste samples were first cast to identify hydration products using X‐ray diffraction, then observed at early age using confocal laser scanning microscopy (CLSM) and at early and late age using scanning electron microscopy. In addition, concrete cores impregnated with fluorescein in order to highlight porosity, cracking and aggregates debonding were observed under UV light using optical microscopy (OM), showing a complete absence of cracking and aggregate debonding. Both microscopy techniques (CLSM and UV light OM) have been applied to this type of binder for the first time. The results show that SSC microstructure is characterised by a sophisticated intergrowth of various phases, including ettringite and amorphous calcium‐(alumina)‐silicate hydrate gels. Finally, Monte–Carlo simulation of electron‐matter has been provided for a better understanding of EDS analysis. LAY DESCRIPTION: This work focuses on the study of a particular construction material known as super sulphated cement (SSC). SSC appears to be a promising alternative to traditional building materials as it offers improved durability reduced and environmental impact attributed to its composition comprising at least 75% of a by‐product of the steel industry. This study investigates the microstructure and properties of this type of cement, focusing particularly on its complex microstructure and the challenges posed by the analysis of its hydration products. To do this, samples of SSC cement paste were first cast to identify hydration products using X‐ray diffraction. They were then examined at early stages using confocal laser scanning microscopy (CLSM) and at early and late stages using scanning electron microscopy (SEM). In addition, fluorescein‐impregnated SSC concrete cores were observed under ultraviolet (UV) light using optical microscopy (OM) to highlight porosity, cracking and delamination of aggregates, phenomena that may be present in such materials. These observations revealed the absence of these problems on this type of cement in this work. This research marks the first application of CLSM and OM under UV light to this type of cement. These microscopic techniques enabled us to understand the hydration phenomenon of this type of cement and its highly complex morphology. Finally, Monte–Carlo simulation was used in this study to model electron‐matter interaction, enabling us to understand the elemental analyses obtained and to gain a better understanding of the microstructure of SSC samples. [ABSTRACT FROM AUTHOR]

In this research, we explore the synthesis of and characterize α-aminophosphonates derived from anthraquinone and benzanthrone, focusing on their fluorescence properties and potential applications in confocal laser scanning microscopy (CLSM). The synthesized compounds exhibit notable solvatochromic behavior, emitting fluorescence from green to red across various solvents. Spectroscopic analysis, including 1H-, 13C-, and 31P-NMR, FTIR, and mass spectrometry, confirms the chemical structures. The compounds' toxicity is evaluated using etiolated wheat sprouts, revealing varying degrees of impact on growth and oxidative damage. Furthermore, the study introduces these α-aminophosphonates for CLSM imaging of the parasitic flatworm Opisthorchis felineus, demonstrating their potential in visualizing biological specimens. Additionally, an X-ray crystallographic study of an anthraquinone α-aminophosphonate provides valuable structural insights. [ABSTRACT FROM AUTHOR]

This paper introduces an innovative microscopy analysis methodology to preserve in situ non-soluble particles within hailstones using a protective porous plastic coating, overcoming previous limitations related to melting the hailstone sample. The method is composed of two techniques: trapping non-soluble particles beneath a plastic coat by using the adapted sublimation technique and then analyzing the particles individually with both Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy with Energy-Dispersive Spectroscopy (SEM-EDS). CLSM provides insights into physical attributes like particle size and surface topography, enhancing understanding of ice nucleation. SEM-EDS complements CLSM by offering detailed information on individual particle elemental chemistry, enabling classification based on composition. Strategies to reduce background noise from glass substrates during EDS spectral analysis are proposed. By combining powerful, high resolution microscopy techniques, this methodology provides valuable data on hailstone composition and properties. This information can give insights into hail developmental processes by enhancing our understanding of the role of atmospheric particles. [ABSTRACT FROM AUTHOR]

A comprehensive in‐situ analysis of the developing gluten network during kneading is still a gap in cereal science. With an in‐line microscale shear kneading and measuring setup in a conventional rheometer, a first step was taken in previous works toward fully comprehensible gluten network development evaluation. In this work, this setup was extended by an in‐situ optical analysis of the evolving gluten network. By connecting a laser scanning microscope with a conventional rheometer, the evaluation of the rheological and optical protein network evolution was possible. An image processing tool for analyzing the protein network was applied for evaluating the gluten network development in a wheat dough during the shear kneading process. This network evaluation was possible without interruption or invasive sample transfer comparing it to former approaches. The shear kneading system was able to produce a fully developed dough matrix within 125% of the reference dough development time in a classical kneader. The calculated network connectivity values from frequency testing ranged over all samples was in good agreement with traditional kneaded wheat dough just over peak consistency. [ABSTRACT FROM AUTHOR]

Background and Aims During the analysis of plant male meiocytes coming from destroyed meiocyte columns (united multicellular structures formed by male meiocytes in each anther locule), a considerable amount of information becomes unavailable. Therefore, in this study intact meiocyte columns were studied by volume microscopy in wild-type rye for the most relevant presentation of 3-D structure of rye meiocytes throughout meiosis. Methods We used two types of volume light microscopy: confocal laser scanning microscopy and non-confocal bright-field scanning microscopy combined with alcohol and aldehyde fixation, as well as serial block-face scanning electron microscopy. Key Results Unusual structures, called nuclear protuberances, were detected. At certain meiotic stages, nuclei formed protuberances that crossed the cell wall through intercellular channels and extended into the cytoplasm of neighbouring cells, while all other aspects of cell structure appeared to be normal. This phenomenon of intercellular nuclear migration (INM) was detected in most meiocytes at leptotene/zygotene. No cases of micronucleus formation or appearance of binucleated meiocytes were noticed. There were instances of direct contact between two nuclei during INM. No influence of fixation or of mechanical impact on the induction of INM was detected. Conclusions Intercellular nuclear migration in rye may be a programmed process (a normal part of rye male meiosis) or a tricky artefact that cannot be avoided in any way no matter which approach to meiocyte imaging is used. In both cases, INM seems to be an obligatory phenomenon that has previously been hidden by limitations of common microscopic techniques and by 2-D perception of plant male meiocytes. Intercellular nuclear migration cannot be ignored in any studies involving manipulations of rye anthers. [ABSTRACT FROM AUTHOR]

Conjugated oligoelectrolytes (COEs) comprise a class of cell‐membrane intercalating molecules that serve as effective optical reporters. However, little is known about the photophysical properties of COEs in biological environments such as buffers, cell membranes, and intracellular organelles, which is critical to optimize performance. Herein, how COE self‐assembly depends on the dielectric environment (polarity and ion content) is explored based on the representative molecule 6‐ring phenylenevinylene (PV) conjugated oligoelectrolyte (COE‐S6), and its optical properties within mammalian cells are subsequently studied. Two‐photon fluorescence lifetime imaging microscopy (FLIM), confocal laser scanning microscopy, and optical properties in solutions are brought together to obtain information about the location, accumulation, and characteristics of the local surroundings. FLIM imaging lifetime phasor plots, decays, and fluorescence spectra on stained mammalian cells provide evidence of successful COE‐S6 internalization via endocytosis. The fluorescence lifetime of COE‐S6 is identical when in A549 mammalian cells and in giant unilamellar vesicle model membranes, thereby providing a correlation between living system and artificial constructs. [ABSTRACT FROM AUTHOR]

The properties and functions of non-covalent interaction-driven fluorescent supramolecular self-assembly depend greatly on their evolution dynamics. Electron microscopy, atomic force microscopy, and confocal laser scanning microscopy have been used to elucidate the formation of molecular self-assembly. However, some pertinent issues, such as the drying or freezing of the sample for electron microscopy, the influence of the interactions between the tip and the sample in atomic force microscopy imaging, and the low spatial resolution of confocal laser scanning microscopy images, often impede the real-time analysis and exploration of the dynamics of molecular self-assembly processes. In this context, fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy have recently been explored to unravel the physical picture of the in situ growth dynamics and stimuli-induced morphological transformation of luminescent self-assembled structures. The current highlight article demonstrates the need for fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy to acquire precise information on the dynamics and morphological evolution of fluorescent self-assembled architectures using a few remarkable recent studies. In addition to the current status and challenges, the future directions for the further exploration of dynamic self-assembly processes towards developing next-generation functional materials have been delineated. [ABSTRACT FROM AUTHOR]

The two fungal human pathogens, Candida auris and Candida albicans, possess a variety of virulence mechanisms. Among them are the formation of biofilms to protect yeast against harsh conditions through the development of (pseudo)hyphae whilst also facilitating the invasion of host tissues. In recent years, increased rates of antifungal resistance have been associated with C. albicans and C. auris, posing a significant challenge for the effective treatment of fungal infections. In the course of our ongoing search for novel anti-infectives, six selected azaphilones were tested for their cytotoxicity and antimicrobial effects as well as for their inhibitory activity against biofilm and hyphal formation. This study revealed that rubiginosin C, derived from stromata of the ascomycete Hypoxylon rubiginosum, effectively inhibited the formation of biofilms, pseudohyphae, and hyphae in both C. auris and C. albicans without lethal effects. Crystal violet staining assays were utilized to assess the inhibition of biofilm formation, while complementary microscopic techniques, such as confocal laser scanning microscopy, scanning electron microscopy, and optical microscopy, were used to investigate the underlying mechanisms. Rubiginosin C is one of the few substances known to effectively target both biofilm formation and the yeast-to-hyphae transition of C. albicans and C. auris within a concentration range not affecting host cells, making it a promising candidate for therapeutic intervention in the future. [ABSTRACT FROM AUTHOR]

Adult isopods of the family Dajidae are exclusively ectoparasitic, typically infecting pelagic malacostracan crustaceans. It is assumed that their life cycle involves free-living and parasitic phases, with planktonic copepods acting as intermediate hosts. Most generic diagnoses proposed in the family have traditionally been incomplete, containing imperfect or misleading information, and characters whose states were wrongly assessed. In an attempt to analyse this state of affairs comprehensively, a taxonomic review and updated diagnosis of the species-rich genus Holophryxus are presented. Both traditional and novel morphological characters are critically assessed, forming the basis for updated differential diagnoses of all currently valid species. The presence of previously ignored sensory structures on the body and pereopods is highlighted and its significance discussed. Holophryxus citriformis sp. nov. is recorded on the common shrimp, Hymenodora glacialis , at 4300 m depth in the central Arctic Ocean. Its description, based on light microscopy and confocal laser scanning microscopy analysis of the female holotype, containing a dwarf male inside its marsupium, is proposed as a model of enhanced descriptive standards required in future morphological research on epicaridean isopods. Definitive host records and geographical distributions of all Holophryxus species are summarized as well as records of larval epicaridean stages associated with copepod hosts. [ABSTRACT FROM AUTHOR]

• Can pigments within a paint hinder proteinous binder characterization using immunofluorescence microscopy?. • Eight out of 10 tested pigments did not interfere with binder characterization using immunofluorescence microscopy. • Five pigments exhibited weak autofluorescence, which did not interfere with immunofluorescence microscopy. • Confocal fluorescence gave clearer results to widefield fluorescence. • Cross sections with smooth surfaces gave a much clearer immunofluorescence microscopy picture. The aim of the study was to address the problematics of proteinous binder characterization, within a cross section of painted model samples, using immunofluorescence microscopy. Problems arise from certain pigments which can alter the epitope sites of target binders (lead white and verdigris) or can exhibit a strong natural autofluorescence (lake pigments). Therefore, dual layered model samples were prepared containing a lower egg tempera paint and an upper oil paint, and both paints were made of the same pigment. As an extra challenge for fluorescence microscopy, half of samples were additionally covered with pure linseed oil (as a third layer), which is known to physically reflect fluorescence. Cross-sections were hybridized with anti-ovalbumin antibodies and with FITC labelled secondary antibodies. To reduce unspecific fluorescence, apart from widefield fluorescence, laser-scanning confocal immunofluorescence microscopy was performed. Finally, 3D surface topography models were constructed which were used to check off any unspecific fluorescence originating from cracks or holes. Results show that immunofluorescence microscopy in the widefield observation mode was successful in its specificity and clarity of highlighting the egg paint layer in the presence of 8 out of 10 pigments, including the problematic lead white and verdigris pigments. Several of these pigments (lead white, malachite, yellow ochre, madder lake and carbon black) exhibited autofluorescence; however it was not bright enough to interfere with the successful immunofluorescence microscopy result. In the widefield mode, immunofluorescence microscopy was unsuccessful in the presence of 2 pigments; carmine lake (pigment adsorbs antibodies) and Dragon's blood (pigment dissolved during resin curing at 50 °C). The confocal observation mode in comparison to the widefield mode achieved a much more specific and clear immunofluorescence microscopy picture (especially in the presence of Prussian blue, Vermilion and carbon black) and removed nearly all of the unspecific fluorescence originating from resin's surface reflection, from pure oil binder, from small indentations and from illumination glair that would have otherwise spread across different layers. Lastly, 3D topography models showed that in general samples with smooth surfaces gave a much clearer immunofluorescence microscopy picture. [ABSTRACT FROM AUTHOR]

The muscid genera Alluaudinella Giglio‐Tos, 1895, Aethiopomyia Malloch, 1921 and Ochromusca Malloch, 1927 form a monophyletic group supported by immature and adult morphology and a highly specialised snail‐feeding strategy of immature stages. In contrast to the undoubted monophyly of the Alluaudinella‐Aethiopomyia‐Ochromusca clade, previous studies have provided contradictory hypotheses of the subfamiliar position within the Muscidae, and these three genera have been placed in the subfamily Muscinae, Dichaetomyiinae, Phaoniinae and Reinwardtiinae. The systematic position of Alluaudinella, as a representative of Alluaudinella, Aethiopomyia and Ochromusca group, is revised by means of larval morphology, biology and molecular data. Light microscopy (LM), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) are used to study the egg and all larval instars of Alluaudinella flavicornis (Macquart, 1855) and a multilocus Sanger sequencing (mS‐seq) approach to examine position within Muscidae. Results are inconsistent with the traditional, morphology‐based concept of the Alluaudinella‐Aethiopomyia‐Ochromusca clade as closely related to Dichaetomyia Malloch, 1921, and the phylogenetic analysis revealed no support for inclusion within subfamily Phaoniinae. Larval morphology in Alluaudinella differs significantly from that of Dichaetomyia (and other Phaoniinae), but resembles that of genera nested in Reinwardtiinae. Based on molecular data and larval morphology a transfer of Alluaudinella, Aethiopomyia and Ochromusca to the subfamily Reinwardtiinae is proposed. [ABSTRACT FROM AUTHOR]

The presence of mechanoreceptors in glabrous skin allows humans to discriminate textures by touch. The amount and distribution of these receptors defines our tactile sensitivity and can be affected by diseases such as diabetes, HIV-related pathologies, and hereditary neuropathies. The quantification of mechanoreceptors as clinical markers by biopsy is an invasive method of diagnosis. We report the localization and quantification of Meissner corpuscles in glabrous skin using in vivo, non-invasive optical microscopy techniques. Our approach is supported by the discovery of epidermal protrusions which are co-localized with Meissner corpuscles. Index fingers, small fingers, and tenar palm regions of ten participants were imaged by optical coherence tomography (OCT) and laser scan microscopy (LSM) to determine the thickness of the stratum corneum and epidermis and to count the Meissner corpuscles. We discovered that regions containing Meissner corpuscles could be easily identified by LSM with an enhanced optical reflectance above the corpuscles, caused by a protrusion of the strongly reflecting epidermis into the stratum corneum with its weak reflectance. We suggest that this local morphology above Meissner corpuscles has a function in tactile perception. [ABSTRACT FROM AUTHOR]

Objectives: Tissue breakdown was assessed by confocal laser scanning microscopy (CLSM) using autofluorescence around implants with ligatures, on a dog hemimandible. Influence of section thickness on the accuracy of histometrical observations was also evaluated, in comparison with thin sections in light microscopy. Material and Methods: Three months after tooth extraction, implants were placed. Two months after abutment placement, ligatures were placed with no plaque control. 11 months post-implantation, the animal was sacrificed. Undecalcified thin (30 µm) sections were cut, stained and evaluated by light microscopy to be used as a reference. Additional sections were performed, so that another pair of unstained thick sections resulted (250–300 µm). Tissue loss was assessed using histomorphometric parameters under CLSM and was compared to the light microscopy reference ones. Results: Morphometry confirmed tissue loss more pronounced on the "thick" and quick sections, when compared to the time-consuming and technique-sensitive "thin" ones. Conclusions: Within the limits of the present study, the adequacy of histometrical observations under CLSM reveal commensurable information about soft-tissue-bone-implant details, when compared to traditional light microscopy histological protocols. The CLSM investigation may seem demanding, yet the richness of data acquired may justify this approach, provided seatbacks caused by improper manipulation of "thick" sections are avoided. [ABSTRACT FROM AUTHOR]

The enigmatic larvae of the Old World genus Passeromyia Rodhain & Villeneuve, 1915 (Diptera: Muscidae) inhabit the nests of birds as saprophages or as haematophagous agents of myiasis among nestlings. Using light microscopy, confocal laser scanning microscopy and scanning electron microscopy, we provide the first morphological descriptions of the first, second and third instar of P. longicornis (Macquart, 1851) (Diptera: Muscidae), the first and third instar of P. indecora (Walker, 1858) (Diptera: Muscidae), and we revise the larval morphology of P. heterochaeta (Villenueve, 1915) (Diptera: Muscidae) and P. steini Pont, 1970 (Diptera: Muscidae). We provide a key to the third instar of examined species (excluding P. steini and P. veitchi Bezzi, 1928 (Diptera: Muscidae)). Examination of the cephaloskeleton revealed paired rod‐like sclerites, named 'rami', between the lateral arms of the intermediate sclerite in the second and third instar larva. We reveal parastomal bars fused apically with the intermediate sclerite, the absence of which has so far been considered as apomorphic for second and third instar muscid larvae. Examination of additional material suggests that modified parastomal bars are not exclusive features of Passeromyia but occur widespread in the Muscidae, and rami may occur widespread in the Cyclorrhapha. [ABSTRACT FROM AUTHOR]

Several species of Passiflora in the subgenus Decaloba bear yellow, circular glands that are between the major veins on the abaxial leaf surface, called ocelli, and commonly described as extrafloral nectaries (EFNs). Although poorly known anatomically, other studies have shown that for some Passiflora species these glands are ecologically important for playing a role in mutualistic relationships with ants. In this study, we determined the nectariferous nature and described the structure of ocelli in the leaves of Passiflora pohlii. We tested the exudates from ocelli for glucose and looked for fungi on the surface of the glands using scanning electron and confocal laser light microscopy. Ocelli of P. pohlii have an epidermis with multiple layers and secretory (or subsecretory) parenchyma. Two ant genera were found collecting profuse nectar from the glands. Fungal hyphae were found at the post-secretory phase. Altogether, these findings confirm that the ocelli of P. pohlii are extrafloral nectaries of ecological importance to the mutualistic interactions between visiting ants and these nectar-secreting structures. [ABSTRACT FROM AUTHOR]

The present work investigates the effect of process parameters on the geometry of wire arc additive manufactured parts. The geometric accuracy of features produced with a weaving strategy is compared to what can be accomplished with a typical overlapping bead strategy. In this work, single-layer and multi-layer geometries were deposited under varying process and path parameters. The wavelength, amplitude, and torch speed of the weaving path were varied, while the power, wire feed speed, and contact tip to work distance remained constant. The geometric deposition efficiencies of several samples produced with a weave strategy are directly compared to samples generated with two parallel overlapping beads with torch speeds defined to match the deposition rate of the weave samples. Feature geometries were characterized using optical microscopy and laser scanning data. The results indicate that implementing a weave path strategy can improve the geometric accuracy of wire arc additive features, thus increasing the effective volumetric deposition rate of the process. It is shown that the most consistent improvements resulted from the combination of low wavelength and high amplitude, which correlate to wider and taller printed layers. [ABSTRACT FROM AUTHOR]

Objective: To explore the mechanism of ropivacaine promoting apoptosis of hepatoma carcinoma cell based on mitochondrial dysfunction and caspase-3 signaling pathway. Methods: Human hepatoma carcinoma cell line BEL-7402 was used for experimental study. After Bel-7402 cells were treated with different concentrations of ropivacaine, the proliferation of hepatoma carcinoma cell was detected by MTT method, the cell morphology was observed by light microscopy and DAPI solution staining, the cell viability was determined by trypan blue staining, the apoptosis of Bel-7402 cells was analyzed by flow cytometry, and the mitochondria were observed under electron microscopy. The nuclear migration of caspase-3 in Bel-7402 cells was observed by laser confocal microscopy, and the effects of ropivacaine on the expression of cytoplasmic apoptosis-related proteins, mitochondrial apoptosis-related proteins, Bel-7402 cells and mitochondrial apoptosis-related proteins were evaluated by western blot assay. Results: Ropivacaine can inhibit the growth of hepatoma carcinoma cell in a dose-dependent and time-dependent manner. The apoptosis of Bel-7402 cells was induced by ropivacaine, and the apoptosis rate of Bel-7402 cells was significantly increased. Ropivacaine can injure mitochondrial function of hepatoma carcinoma cells. Laser confocal microscopy showed the migration of caspase-3 molecules to the nucleus. Ropivacaine interacts with caspase-3 to promote the migration of caspase-3 into the nucleus, stimulate the expression of caspase-3, PARP-1 and caspase-9 proteins, inhibit the expression of Bcl-2, promote the expression of APAF-1, promote mitochondria to release Cytochrome C and activate caspase-3 activity. Conclusion: Ropivacaine can promote the apoptosis of hepatoma carcinoma cell, and its mechanism may be related to the destruction of mitochondrial function and activation of caspase-3 signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To explore the mechanism of ropivacaine promoting apoptosis of hepatoma carcinoma cell based on mitochondrial dysfunction and caspase-3 signaling pathway. Methods: Human hepatoma carcinoma cell line BEL-7402 was used for experimental study. After Bel-7402 cells were treated with different concentrations of ropivacaine, the proliferation of hepatoma carcinoma cell was detected by MTT method, the cell morphology was observed by light microscopy and DAPI solution staining, the cell viability was determined by trypan blue staining, the apoptosis of Bel-7402 cells was analyzed by flow cytometry, and the mitochondria were observed under electron microscopy. The nuclear migration of caspase-3 in Bel-7402 cells was observed by laser confocal microscopy, and the effects of ropivacaine on the expression of cytoplasmic apoptosis-related proteins, mitochondrial apoptosis-related proteins, Bel-7402 cells and mitochondrial apoptosis-related proteins were evaluated by western blot assay. Results: Ropivacaine can inhibit the growth of hepatoma carcinoma cell in a dose-dependent and time-dependent manner. The apoptosis of Bel-7402 cells was induced by ropivacaine, and the apoptosis rate of Bel-7402 cells was significantly increased. Ropivacaine can injure mitochondrial function of hepatoma carcinoma cells. Laser confocal microscopy showed the migration of caspase-3 molecules to the nucleus.Ropivacaine interacts with caspase-3 to promote the migration of caspase-3 into the nucleus, stimulate the expression of caspase-3, PARP-1 and caspase-9 proteins, inhibit the expression of Bcl-2, promote the expression of APAF-1, promote mitochondria to release Cytochrome C and activate caspase-3 activity. Conclusion: Ropivacaine can promote the apoptosis of hepatoma carcinoma cell, and its mechanism may be related to the destruction of mitochondrial function and activation of caspase-3 signaling pathway. [ABSTRACT FROM AUTHOR]

Advances in microscopy hardware and storage capabilities lead to increasingly larger multidimensional datasets. The multiple dimensions are commonly associated with space, time, and color channels. Since "seeing is believing", it is important to have easy access to user-friendly visualization software. Here we present IMAGE-IN, an interactive web-based multidimensional (N-D) viewer designed specifically for confocal laser scanning microscopy (CLSM) and focused ion beam scanning electron microscopy (FIB-SEM) data, with the goal of assisting biologists in their visualization and analysis tasks and promoting digital workflows. This new visualization platform includes intuitive multidimensional opacity fine-tuning, shading on/off, multiple blending modes for volume viewers, and the ability to handle multichannel volumetric data in volume and surface views. The software accepts a sequence of image files or stacked 3D images as input and offers a variety of viewing options ranging from 3D volume/surface rendering to multiplanar reconstruction approaches. We evaluate the performance by comparing the loading and rendering timings of a heterogeneous dataset of multichannel CLSM and FIB-SEM images on two devices with installed graphic cards, as well as comparing rendered image quality between ClearVolume (the ImageJ open-source desktop viewer), Napari (the Python desktop viewer), Imaris (the closed-source desktop viewer), and our proposed IMAGE-IN web viewer. [ABSTRACT FROM AUTHOR]

Ursolic acid (UA) exists in a variety of medicinal plants. UA exhibits antimicrobial activity against several microorganisms; however, little is known regarding the potential antifungal effect of UA on Cryptococcus neoformans (C. neoformans). The antifungal and antibiofilm activities of UA on C. neoformans H99 were evaluated in this study. Minimum inhibitory concentration (MIC) of UA against C. neoformans H99 was determined by microdilution technique, and its action mode was elucidated by clarifying the variations in cell membrane integrity, capsule, and melanin production. Moreover, the inhibition and dispersal effects of UA on biofilm formation and mature biofilms by C. neoformans H99 were evaluated using crystal violet (CV) assay, optical microscopy, field emission scanning electron microscopy and confocal laser scanning microscopy. The results indicated that the MIC value of UA against C. neoformans H99 was 0.25 mg/mL. UA disrupted the cell membrane integrity, inhibited the capsule and melanin production of C. neoformans H99 in a concentration-dependent manner. Further, UA presented the inhibitory effect on biofilm formation and dispersed mature biofilms, as well as compromised the cell membrane integrity of C. neoformans H99 cells within biofilms. Together, these results indicate that UA might be a potential therapeutic option for the treatment of C. neoformans-related infections. [ABSTRACT FROM AUTHOR]

Studying leaves of aquatic and terrestrial populations of Phragmites australis grown on the bank of the Venetian strait of the Dnipro River in Kyiv (Ukraine) for establishing the role of bulliform cells in the mechanisms of plant resistance to moderate soil drought were studied. Bulliform (motor) cells participate in the twisting and folding of leaves during drought and strong sunlight. The study of bulliform cells in the leaves of the aquatic and terrestrial populations of P. australis was carried out using the methods of light microscopy, cytochemical methods, laser confocal microscopy, and biochemical methods. The comparative analysis of the structure of bulliform cells of leaves of P. australis has shown significant differences depending on plant growth location. The differences in the number, size, and area of bulliform cells and also polysaccharide content showed clear phenotypical plasticity. Cytochemical and laser confocal microscopic studies of polysaccharides of cell walls in bulliform cells of aquatic and terrestrial ecotypes of P. australis showed that a decrease in soil moisture in a natural moderate drought of soil led to an increase in lignin and syringyl monolignol content in the outer walls of bulliform cells and also to a decrease in cellulose and callose content in outer and inner cell walls. The obtained data shows that the studied signs of bulliform cells can be markers of tolerance for population plants that have the ability to curl leaves for the preservation of optimal water balance in moderate drought. [ABSTRACT FROM AUTHOR]

Stimulated emission depletion (STED) microscopy achieves super-resolution by exciting a diffraction-limited volume and then suppressing fluorescence in its outer parts by depletion. Multiple depletion lasers may introduce misalignment and bleaching. Hence, a single depletion wavelength is preferable for multi-color analyses. However, this limits the number of usable spectral channels. Using cultured cells, common staining protocols, and commercially available fluorochromes and microscopes we exploit that the number of fluorochromes in STED or confocal microscopy can be increased by phasor based fluorescence lifetime separation of two dyes with similar emission spectra but different fluorescent lifetimes. In our multi-color FLIM-STED approach two fluorochromes in the near red (exc. 594 nm, em. 600–630) and two in the far red channel (633/641–680), supplemented by a single further redshifted fluorochrome (670/701–750) were all depleted with a single laser at 775 nm thus avoiding potential alignment issues. Generally, this approach doubles the number of fully distinguishable colors in laser scanning microscopy. We provide evidence that eight color FLIM-STED with a single depletion laser would be possible if suitable fluorochromes were identified and we confirm that a fluorochrome may have different lifetimes depending on the molecules to which it is coupled. [ABSTRACT FROM AUTHOR]

Objective Study to establish an in vitro method for the isolation, culture and identification of endothelial cells from the penile corpus cavernosum of SD rats. Methods The endothelial cells were purified by collagenase digestion and mechanical curettage. After 11 days of culture in EGM-2 complete culture medium, the cells were subcultured for the second generation. After hematoxylin-eosin staining, the endothelial cells were observed under light microscope, and the cell growth curve was drawn by CCK8 method. The expression of CD31 and vWF in endothelial cells was identified by immunofluorescence method, the cycle changes of endothelial cells were detected by flow cytometry, and the expression changes of vWF marker in endothelial cells were observed by laser confocal microscopy. Results The endothelial cells of rat penile corpus cavernosum isolated by collagenase digestion covered more than 2/3 of the culture flask area in 11 ~14d. After HE staining, the cells showed typical paving stone shape under light microscope. The co-positive expression rate of CD31 and vWF was (86. 19±1. 14)%. VWF could be stably expressed in the first 4 generations of cells and the growth cycle of the second and third generations of cells was stable. After hematoxylin-eosin staining, the cells appear typical pavement-like under light microscopy. Conclusion High purity endothelial cells from rat penile corpus cavernosum can be successfully isolated by collagenase digestion combined with mechanical scraping,which can be used as a cellular model for the study of endothelial cells in the penile corpus cavernosum. [ABSTRACT FROM AUTHOR]

Microscopy imaging of living cells is becoming a pivotal, noninvasive, and highly specific tool in metabolic engineering to visualize molecular dynamics in industrial microorganisms. This review describes the different microscopy methods, from fluorescence to super resolution, with application in microbial bioengineering. Firstly, the role and importance of microscopy imaging is analyzed in the context of strain design. Then, the advantages and disadvantages of different microscopy technologies are discussed, including confocal laser scanning microscopy (CLSM), spatial light interference microscopy (SLIM), and super-resolution microscopy, followed by their applications in synthetic biology. Finally, the future perspectives of live-cell imaging and their potential to transform microbial systems are analyzed. This review provides theoretical guidance and highlights the importance of microscopy in understanding and engineering microbial metabolism. In the construction of cell factories, microscopy imaging of living cells plays an important role in the 'build' and 'test' of the 'design–build–test–learn' cycle. Confocal laser scanning microscopy and its extended technologies can be used to study the regulation of microbial metabolism, enzyme assembly, detection of enzyme expressing level, and localization analysis of transporters and other membrane proteins. Variable-angle total internal reflection fluorescence microscopy has been gradually applied to reveal heterogeneity or physical effects that are usually hidden in the average measures of bacteria populations. The remarkable advantages of super-resolution imaging technologies for the identification of single-molecules with nanometer resolution will further provide novel insights in metabolic engineering and synthetic biology. [ABSTRACT FROM AUTHOR]

The forewing base structure of the orthopteran suborder Ensifera, a group of insects well known for their sound-producing behavior, was examined by using light microscopy, confocal laser scanning microscopy, and µCT. We detected significant novel characters, such as functional changes in the tpm9 muscle from the flexor to the extensor of the forewing that are likely associated with the acquisition of sound-producing behavior using forewings. Phylogenetic analysis based on the characters selected from the forewing base structure showed that the character system contains a strong phylogenetic signal supporting the monophyly of Ensifera, Tettigonioidea and Grylloidea as well as the sister-group relationship between Grylloidea and Gryllotalpoidea, but no apomorphies to resolve the deep phylogeny among superfamilies could be determined from this character system. [ABSTRACT FROM AUTHOR]

The effect of lactocin AL705, bacteriocin produced by Latilactobacillus (Lat.) curvatus CRL1579 against Listeria biofilms on stainless steel (SS) and polytetrafluoroethylene (PTFE) coupons at 10 °C was investigated. L. monocytogenes FBUNT showed the greatest adhesion on both surfaces associated to the hydrophobicity of cell surface. Partially purified bacteriocin (800 UA/mL) effectively inhibited L. monocytogenes preformed biofilm through displacement strategy, reducing the pathogen by 5.54 ± 0.26 and 4.74 ± 0.05 log cycles at 3 and 6 days, respectively. The bacteriocin-producer decreased the pathogen biofilm by ∼2.84 log cycles. Control and Bac− treated samples reached cell counts of 7.05 ± 0.18 and 6.79 ± 0.06 log CFU/cm2 after 6 days of incubation. Confocal scanning laser microscopy (CLSM) allowed visualizing the inhibitory effect of lactocin AL705 on L. monocytogenes preformed biofilms under static and hydrodynamic flow conditions. A greater effect of the bacteriocin was found at 3 days independently of the surface matrix and pathogen growth conditions at 10 °C. As a more realistic approach, biofilm displacement strategy under continuous flow conditions showed a significant loss of biomass, mean thickness and substratum coverage of pathogen biofilm. These findings highlight the anti-biofilm capacity of lactocin AL705 and their potential application in food industries. • Lactocin AL705 inhibited L. monocytogenes biofilm through displacement strategy. • The greatest effectiveness was found on SS and PTFE surfaces after 3 days at 10 °C. • Bacteriocin reduced of biofilm biomass and thickness in continuous flow conditions. • Lactocin AL705 proved to be an effective and promising anti-biofilm agent. [ABSTRACT FROM AUTHOR]

Understanding how late an inhibitor can be released once corrosion initiated without compromising corrosion protection may help in developing more efficient anticorrosion coatings. We explored this idea through time-controlled Ce(NO 3) 3 availability to AA2024-T3 immersed in 0.05 M NaCl. Ce(NO 3) 3 was supplied at 0, 30, 60, and 180 s from the start of immersion to get a concentration of 0.001 M. Detailed visualization of surface changes at the intermetallic particle level was obtained using in-situ reflected microscopy. SEM-EDX and confocal laser microscopy confirmed the extent of intermetallic degradation and local inhibitor deposition corresponding to operando changes. When the inhibitor is supplied within 60 s of immersion, the surface changes slowdown earlier and are visually less extensive than in uninhibited systems. Furthermore, our results highlight the potential of reflected microscopy for local corrosion inhibition studies and underscore the importance of understanding the interaction between inhibitor release timing and corrosion protection. • A reflected microscopy-based protocol for detailed visualization of local inhibitor deposition presented. • An analysis protocol of pixels activity is proposed to distinguish trenching and dealloying from inhibitor deposition. • Letting intermetallic particles dealloy for a brief period significantly affects Ce(III) capacity to precipitate. • Corrosion protection is not compromised when Ce(III) is supplied to the IMPs in our model system within 60 s of exposure. [ABSTRACT FROM AUTHOR]

Skin ageing is a complex phenomenon linked to intrinsic and extrinsic factors, such as ultraviolet exposure, smoking or pollution,1-4 which involves all skin layers.5-7 Ageing is characterized by a reduction in collagen synthesis,4,8,9 a decreased hydration,5,6 epidermal thickness variations,7 and an accelerated breakdown and disorganization of collagen fibres4,8,9 as well as loss of skin elasticity.10 Chemical peelings can potentially help to reverse these degenerative processes. In accordance with skin imaging results, 80% of patients ( I N i = 12) were very satisfied, reporting an improved skin texture, a plumped appearance in the major part of cases (73.3%) and a brighter face, which made it unnecessary to apply routine make-up. [Extracted from the article]

The objectives of this work were to image the liposarcoma with light microscopy and measure the absorbance spectra of liposarcoma and different mammalian blood in range of ultraviolet (UV), visible region and infrared (IR) range. We have made the systematic analysis of mammalian blood of species with a high (human), medium (cow) and low (goat) of absorbance. A customized optical diode laser light source microscope (laser wavelength, λo=630 nm and bandwidth, Δλ=15 nm) and an UV/visible spectrophotometer (wavelength, λ=190 to 1100 nm) have been used for investigation of well differentiated liposarcoma in abdomen tissues and mammalian blood. We have observed that the shapes and size of the cells are not uniform throughout the cancerous tissue. The marks on the cells and dead cells were also there in plenty. Cellular spaces between the cells have also been observed. We have seen that the shapes and size of the cells shows unpredictability. [ABSTRACT FROM AUTHOR]

A Tikhonov-regularized solution of the inverse scattering problem has been physically implemented using a basis of speckles that is adjusted immediately during an experiment and ensures the minimization of regularization-induced diffusion of images. This procedure ensures the stable, minimally diffuse reconstruction of images in the scheme of microscopy of scattering media with a submicron spatial resolution. [ABSTRACT FROM AUTHOR]

Summary: The objective of this work was to extract high methoxyl (HM) pectin with oxalic acid, characterise its physicochemical properties and evaluate its dispersion stability in acidified milk drink (AMD). HPSEC–MALLS analysis revealed that the obtained HM pectin was high in molecular weight (522.4 kDa). The ζ‐potential of the obtained HM pectin in aqueous solution at pH 4.0 was −31.5 mV. The AFM images proved that the obtained HM pectin contained several long linear molecules. The apparent diameter of AMD stabilised by the obtained HM pectin was 636 nm at the concentration of 3.0‰. The stability analysis of AMD and the microscopic observation with confocal laser scanning microscopy (CLSM) proved that the obtained HM pectin was a prominent stabiliser in AMD. [ABSTRACT FROM AUTHOR]

Titanium has a good corrosion resistance, owing to a thin, compact oxide layer that is formed spontaneously when the metal is exposed to the environment. However, even titanium can be subject to corrosion, for example, when exposed to reducing strong acids. This paper aims to analyze the differences in corrosion resistance of commercially pure titanium (Ti grade 2) and the most resistant Ti–Pd alloy (Ti grade 7, ∼0.2% Pd) in a strong reducing acid environment. Immersion tests, with results collected in the form of corrosion rates, are provided in the first part of the present work. Corrosion morphology is investigated using optical microscopy and laser profilometry. Then, an electrochemical characterization of both metals has been performed by means of cathodic polarization tests, electrochemical impedance spectroscopy, and linear polarization resistance. Weight loss data, electrochemical tests, and corrosion morphology are in good agreement: H2SO4, HCl, and oxalic acid are the most aggressive chemicals for commercially pure titanium. In such conditions, the use of Ti grade 7 appears mandatory. [ABSTRACT FROM AUTHOR]

Studies of female genital structures have generally lagged behind comparable studies of male genitalia, in part because of an assumption of a lower level of variability, but also because internal genitalia are much more difficult to study. Using multiple microscopy techniques, including video stereomicroscopy, fluorescence microscopy, low-temperature scanning electron microscopy (LT-SEM), and confocal laser scanning microscopy (CLSM) we examined whether the complex sperm transfer structures in males of Megalolaelaps colossus (Acari: Mesostigmata) are matched by similarly complex internal structures in the female. While both LT-SEM and CLSM are well suited for obtaining high-quality surface images, CLSM also proved to be a valuable technique for observing internal anatomical structures. The long and coiled sperm transfer organ on the chelicera of the males (spermatodactyl) largely matches an equally complex, but internal, spiral structure in the females in shape, size, and direction. This result strongly suggests some form of genital coevolution. A hypothesis of sexual conflict appears to provide the best fit for all available data (morphology and life history). [ABSTRACT FROM AUTHOR]

We describe new insights into the morphology and life history of the bizarre parasite Haplozoon axiothellae (Dinoflagellata) using light microscopy (LM), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Trophonts were isolated from the intestines of host maldanid polychaetes, Axiothella rubrocincta, collected from San Juan Island, Washington, USA. LM and SEM confirmed features previously observed, such as amphiesmal projections, mature and immature junctions between the nucleated compartments of the vermiform syncytium and visible polygonal alveoli. CLSM of adult trophonts fluorescently stained for DNA, tubulin, centrin, and plasma membrane demonstrated several new ultrastructural traits: (1) an extensive basket of parallel microtubules within the trophomere used for host attachment, (2) two physically separated MTOCs (i.e. putative pairs of basal bodies) beneath pores on the ventral side of each compartment, (3) robust mitotic and/or meiotic spindles associated with one to four nuclei in each compartment, (4) spindles with polar bodies that are disconnected from the MTOCs, (5) a centrin-stained fibril within the trophomeres that potentially functions to retract the motile stylet, and (6) cytokinesis in the posterior-most compartments. This study renames haplozoan compartments using the suffix "-mere" rather than "-cyte" (i.e. trophomere, gonomere, sporomere) to reflect their status within a single syncytium. [ABSTRACT FROM AUTHOR]

Fluorescence-based microscopy as one of the standard tools in biomedical research benefits more and more from super-resolution methods, which offer enhanced spatial resolution allowing insights into new biological processes. A typical drawback of using these methods is the need for new, complex optical set-ups. This becomes even more significant when using two-photon fluorescence excitation, which offers deep tissue imaging and excellent z-sectioning. We show that the generation of striped-illumination patterns in two-photon laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement using open-source image reconstruction software. The special appeal of this approach is that even in the case of a commercial two-photon laser scanning microscope no optomechanical modifications are required to achieve this modality. Modifying the scanning software with a custom-written macro to address the scanning mirrors in combination with rapid intensity switching by an electro-optic modulator is sufficient to accomplish the acquisition of two-photon striped-illumination patterns on an sCMOS camera. We demonstrate and analyse the resulting resolution improvement by applying different recently published image resolution evaluation procedures to the reconstructed filtered widefield and super-resolved images. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 1)'. [ABSTRACT FROM AUTHOR]

In this work, nanocomposites that combine mucopenetrating and mucoadhesive properties in a single system are proposed as innovative strategy to increase drug residence time in the intestine following oral administration. To this aim, novel mucoadhesive chitosan (CH) sponges loaded with mucopenetrating nanoemulsions (NE) were developed via freeze-casting technique. The NE mucopenetration ability was determined studying the surface affinity and thermodynamic binding of the nanosystem with mucins. The ability of nanoparticles to penetrate across a preformed mucins layer was validated by 3D-time laps Confocal Laser Scanning Microscopy imaging. Microscopy observations (Scanning Electron Microscopy and Optical Microscopy) showed that NE participated in the structure of the sponge affecting its stability and in vitro release kinetics. When incubated with HCT 116 and Caco-2 cell lines, the NE proved to be cytocompatible over a wide concentration range. Finally, the in vivo biodistribution of the nanocomposite was evaluated after oral gavage in healthy mice. The intestinal retention of NE was highly enhanced when loaded in the sponge compared to the NE suspension. Overall, our results demonstrated that the developed nanocomposite sponge is a promising system for sustained drug intestinal delivery. [Display omitted] • Nanocomposites embedding nanoemulsions (NE) in chitosan (CH) sponges are produced by freeze-drying. • Modulation of nanocomposite structural features affects sponge stability and NE release in simulated intestinal fluids. • PEGylated NE shows effective mucopenetrating properties. • Mucoadhesive ability of CH sponges increases NE intestinal residence time following oral administration. [ABSTRACT FROM AUTHOR]

The cyclodextrin (CD)-based emulsions exhibit complex instability behaviors such as rapid flocculation and creaming, and how to capture droplet dispersion states of the emulsions remains a great challenge. Here we prepare the CD-based emulsions with different oil-water volume ratios and CD concentrations by using high-pressure homogenization, and characterize the emulsion droplets by using optical microscopy and confocal laser scanning microscopy. We evaluate the effects of homogenization pressure on the stability of the emulsions, identify armored droplets with different surface features, measure interfacial concentrations of adsorbed ICs microcrystals, and observe ejection of the oil/CD inclusion complexes (ICs) microcrystals from the droplet surface. The droplet dispersion states are sensitive to the dynamic buildup and evolving morphologies of the interfacial microcrystals, and there are clear correlations between the properties of the ejected microcrystals and the characteristics of the emulsions. We ascribe the subsequent ejection of ICs microcrystals from the droplet surface to consolidation and deformation of the films formed between neighboring droplets. The ejection of the ICs microcrystals affords a simple method to detect the droplet-droplet interactions and phase transitions in the CD-based emulsions, which might be a generic feature in the broader context of the creaming processes of emulsions. [Display omitted] [ABSTRACT FROM AUTHOR]

This study assessed the gross-structure, micro-structure, physical characteristics and nutritional composition of Trichilia emetica and Trichilia dregeana seeds. T. emetica and T. dregeana seeds have potential for commercialization and improved food security, yet they are under-utilized and under-researched. The gross- and micro-structure of the seeds was assessed using stereo microscopy, light microscopy (LM), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Protein, fat, ash, and mineral content of the seeds were also assessed. T. emetica and T. dregeana seeds had lower bulk density (393.5 kgm−3 & 433.6 kgm−3 respectively) and lower porosity (55.07% & 54.38% respectively) than soybean. Geometric mean diameter (29.7 mm & 16.9 mm) and aspect ratio (1.72 and 1.85) of T. emetica and T. dregeana seeds respectively, were higher than soybean, while they had lower sphericity (0.66 and 0.66 respectively) than soybean (0.99). Microscopy analysis showed that Trichilia species had larger-sized and more round-shaped protein bodies than soybean seeds. T. emetica and T. dregeana protein contents (25.6% and 17.3% w/w, respectively) were lower than soybean (45.4%) while fat contents (49% and 51.5% w/w, respectively) were higher than soybean (20.2% w/w). Potassium (1075–1350 mg100g−1) and calcium (285–300 mg100g−1) where the main macro-minerals while iron (6.33–6.83 mg100g−1) and zinc (2.30–2.90 mg100g−1) were the main micro-minerals. The research demonstrated that the structural and nutritional characteristics of T. dregeana and T. emetica seeds could facilitate their commercial utilization and application for food security alleviation. [ABSTRACT FROM AUTHOR]

The paper aimed to present the microscopic techniques used in botanical origin determination of honey samples, shortly melissopalynology. This technique is used for microscopic examination of pollen grains to determine the botanical origin of honey. It is known that a certain number of pollen grains must be recovered from sediment of honey solution, and the presence of small amounts of pollen may be related to falsification. Identification of pollen structure is generally made using light microscopy; phase contrast microscopy may be also used. Fluorescence microscopy is also a powerful method in palynology. Confocal laser scanning microscopy is effective in revealing the ultrastructure of pollen outer layer and shape of the pollen. The determination of the botanical origin of honey using palynology is based on the relative frequency of the pollen belonging to nectariferous plants. Honey is considered monofloral if the pollen from the sediment comes predominantly from a named botanical origin and overpasses 45% from the total count of pollen grains counted on the microscopic slide. [ABSTRACT FROM AUTHOR]

BACKGROUND Oil‐in‐water (O/W) emulsions are thermodynamically unstable and are easily oxidized. Recently, protein hydrolysates have been used to enhance the emulsifying and oxidative stability of emulsions. High‐pressure processing (HPP) enzymatic hydrolysates of soy protein isolate have higher bioactivities. The objective of the study was to investigate the effects of various soy protein isolate hydrolysate (SPIH) concentrations obtained during different 4 h pressure treatments on improving the emulsifying and oxidative stability of myofibrillar protein (MP) emulsions. RESULTS: Emulsions with 4 mg mL−1 SPIH obtained at 200 MPa had the highest emulsifying activity index and emulsion stability index (P ≤ 0.05). This increase in emulsion stability was related to increased zeta potential and reduced average particle size. Optical microscopy and confocal laser scanning microscopy observations confirmed that emulsions with 4 mg mL−1 SPIH possessed relatively small oil droplets. The addition of SPIH obtained at 200 MPa significantly reduced thiobarbituric acid reactive substance values (P ≤ 0.05) of emulsions during 8 days of storage. Concurrently, the carbonyl content remained the lowest and the sulfhydryl content remained the highest, which indicated that the emulsions had higher protein oxidative stability. CONCLUSIONS: SPIH obtained under HPP could improve the emulsifying and oxidative stability of MP‐prepared O/W emulsions. Highlights: Emulsions were prepared with myofibrillar protein (MP) and soy protein isolate hydrolysates (SPIHs) obtained under various pressures for 4 h.Emulsions with 4 mg mL−1 SPIH obtained at 200 MPa had high emulsion activity and stability.Emulsions with 4 mg mL−1 SPIH obtained at 200 MPa had improved oxidative stability. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The products of secondary xylem are of significant biological and commercial importance, and as a result, the biology of secondary growth and how intrinsic and extrinsic factors influence this process have been the subject of intense investigation. Studies into secondary xylem range in scale from the cellular to the forest stand level, with phenotypic analyses often involving the assessment of traits relating to cell morphology and cell wall chemical composition. While numerous techniques are currently available for phenotypic analyses of samples containing abundant amounts of secondary tissue, only a few of them (microanalytical techniques) are suitable when working with limiting amounts of secondary tissue or where a fine-scale resolution of morphological features or cell wall chemical composition is required. While polarised light microscopy, scanning electron microscopy, field emission-scanning electron microscopy and X-ray scattering and micro-tomography techniques serve as the most frequently used microanalytical techniques in morphotyping, techniques such as scanning ultraviolet microspectrophotometry, X-ray photoelectron spectroscopy, gas chromatography, Fourier-transform infrared spectroscopy and matrix-assisted laser desorption ionisation mass spectrometry serve as the most commonly used microanalytical techniques in chemotyping. Light microscopy, fluorescence microscopy, confocal laser scanning microscopy, transmission electron microscopy and Raman spectroscopy serve as dual micro morphotyping and chemotyping techniques. In this review, we summarise and discuss these techniques in the light of their applicability as microanalytical techniques to study secondary xylem. [ABSTRACT FROM AUTHOR]

Summary: The applicability of confocal laser scanning microscopy is limited, e.g. by attenuation of the excitation and the fluorescence emission beam. As a prerequisite for further processing and analysis of the obtained microscopic images, a new method is presented for correcting this attenuation. The correction is based on beam modelling and on a differential form of the modified Beer–Lambert law. It turns out that the intensity decay can be modelled as a double convolution of the microscopic image with the intensities of the excitation semibeam and the emission beam. Under weak assumptions made for the intensities of the fluorescent radiation and the detected signal, formulas for the attenuation correction and the attenuation simulation are derived. The method traces back to that one published by Roerdink which is modified concerning a more realistic beam modelling, avoiding the so‐called weak attenuation expansion and considering fluorescence excitation throughout the light cone of the excitation beam. The applicability of the method is demonstrated for synthetic examples as well as microscopic images of chromatographic beads. It is shown that the new method can be successfully applied for reconstructing the true fluorophore distribution in specimens even if the microscopic images are affected by strong attenuation. Lay Description: The applicability of confocal laser scanning microscopy is limited by attenuation of the excitation and the fluorescence emission beam. As a prerequisite for further processing and analysis of the obtained microscopic images, a new method is presented for correcting this attenuation. The correction is based on modeling the excitation as well as the emission beam and on a modified Beer‐Lambert law for beam attenuation. The applicability of the method is demonstrated for synthetic examples as well as microscopic images of chromatographic beads. It is shown that the new method can be successfully applied for reconstructing the true fluorophore distribution in specimens even if the microscopic images are affected by strong attenuation. [ABSTRACT FROM AUTHOR]

Aims: Alternate wetting and drying irrigation (AWD) benefits root iron plaque (IP) formation and nutrient uptake, but the underlying mechanism is unclear. Methods: RNA-sequencing, histochemical staining, confocal laser microscopy, and transgenic techniques were used to evaluate the contributions of ionic-bonded cell-wall-located class III peroxidases (iPrxs) and hydrogen peroxide (H2O2) to IP formation in rice (Oryza sativa) under AWD. Results: AWD significantly enhanced iPrx activity and expression of OsPrx24, a gene encoding iPrx, accompanied by elevated H2O2 accumulation and IP formation. Microscopic analysis indicated that OsPrx24 was mainly expressed in root epidermis, where H2O2 accumulation and IP precipitation also occurred. In an OsPrx24-overexpression line (OsPrx24-OX), the H2O2 concentration was 28.2% lower, and IP formation was 42.6% higher than in the wild type (WT). Treatment with both an iPrx inhibitor and an H2O2 scavenger reduced the IP content, but it was higher in OsPrx24-OX than in the WT under these treatment conditions. Phosphorus concentrations were higher and more IP was found in the roots of OsPrx24-OX than the WT. Conclusion: The results imply that OsPrx24 participates in IP formation by regulating iPrx activity and H2O2 concentration under AWD. The results enhance our understanding of IP formation and nutrient uptake in rice. [ABSTRACT FROM AUTHOR]

This study evaluated the organic residues of milk fouling using fluorescence and confocal laser scanning microscopy. The inorganic content was analyzed with energy-dispersive X-ray spectroscopy, complemented with inductively coupled plasma optical emission spectrometry. These techniques were applied to evaluate milk fouling cleanliness using an alkaline product and an enzymatic formulation based on protease and amylase. The results showed that the efficiency of enzymatic cleaning was 87.1% when it was evaluated at 55°C for 30 min, and with a medium of pH 8.5. No difference was found from the efficacy in eliminating dairy fouling observed for the chemical cleaning (86.9%). The fluorescence microscopy proved useful for determining the organic solid components in the outer layer of the dairy fouling. The fouling spatial disposition in 3 dimensions, obtained by confocal laser scanning microscopy, showed that it was formed of 51.3% sugars, 9.3% fats, and 39.4% proteins, with the enzymatic cleaning of these compounds being homogeneous, compared with chemical cleaning. The protein and lipid contents were in the surface layer, whereas sugars were located in the innermost part that contributes to the Maillard reaction during fouling formation. After enzymatic cleaning, the reduction in the concentration of Ca and P was 71.61 and 74.67%, respectively, compared with fouling intact. Thus, enzymatic cleaning, without the accumulation of Na from chemical cleaning, leaves 1.5 times less mineral than chemical cleaning. Knowing the content and structure of fouling in the industry helps to formulate better products to achieve proper levels of cleanliness. Additionally, studying the cleaning residues helps to avoid problems of cross-contamination between batches or subsequent microbial growths (biofilms) on surfaces with residues. [ABSTRACT FROM AUTHOR]

Recent developments in laser scanning microscopy have greatly extended its applicability in cancer imaging beyond the visualization of complex biology, and opened up the possibility of quantitative analysis of inherently dynamic biological processes. However, the physics of image acquisition intrinsically means that image quality is subject to a tradeoff between a number of imaging parameters, including resolution, signal-to-noise ratio, and acquisition speed. We address the problem of geometric distortion, in particular, jaggedness artefacts that are caused by the variable motion of the microscope laser, by using a combination of image processing techniques. Image restoration methods have already shown great potential for post-acquisition image analysis. The performance of our proposed image restoration technique was first quantitatively evaluated using phantom data with different textures, and then qualitatively assessed using in vivo biological imaging data. In both cases, the presented method, comprising a combination of image registration and filtering, is demonstrated to have substantial improvement over state-of-the-art microscopy acquisition methods. [ABSTRACT FROM AUTHOR]

In the present paper, well-preserved specimens of taxonomically distinctive Proterozoic eukaryotic fossil Tappania Yin are recorded for the first time from the rocks of the Saraipali Formation of the Singhora Group, Chhattisgarh Supergroup, India. In the global context, among the various species of this genus, Tappania plana is widely distributed in the latest Palaeoproterozoic (Statherian) to the early Mesoproterozoic (Calymmian) organic-walled microfossil assemblages. Tappania plana of the Saraipali is subjected to transmitted light microscopy, confocal laser scanning microscopy and laser Raman spectroscopy and these results are presented. Collectively, the occurrence of remarkable microfossil Tappania and other associated microfossils in the Saraipalli Formation of rocks demonstrate the Calymmian age for the lower sediments of the Chhattisgarh Supergroup. [ABSTRACT FROM AUTHOR]

Many microbes exhibit quorum sensing (QS) to cooperate, share and perform a social task in unison. Recent studies have shown the emergence of reversible phenotypic heterogeneity in the QS-responding pathogenic microbial population under laboratory conditions as a possible bet-hedging survival strategy. However, very little is known about the dynamics of QS-response and the nature of phenotypic heterogeneity in an actual host-pathogen interaction environment. Here, we investigated the dynamics of QS-response of a Gram-negative phytopathogen Xanthomonas pv. campestris (Xcc) inside its natural host cabbage, that communicate through a fatty acid signal molecule called DSF (diffusible signal factor) for coordination of several social traits including virulence functions. In this study, we engineered a novel DSF responsive whole-cell QS dual-bioreporter to measure the DSF mediated QS-response in Xcc at the single cell level inside its natural host plant in vivo. Employing the dual-bioreporter strain of Xcc, we show that QS non-responsive cells coexist with responsive cells in microcolonies at the early stage of the disease; whereas in the late stages, the QS-response is more homogeneous as the QS non-responders exhibit reduced fitness and are out competed by the wild-type. Furthermore, using the wild-type Xcc and its QS mutants in single and mixed infection studies, we show that QS mutants get benefit to some extend at the early stage of disease and contribute to localized colonization. However, the QS-responding cells contribute to spread along xylem vessel. These results contrast with the earlier studies describing that expected cross-induction and cooperative sharing at high cell density in vivo may lead to synchronize QS-response. Our findings suggest that the transition from heterogeneity to homogeneity in QS-response within a bacterial population contributes to its overall virulence efficiency to cause disease in the host plant under natural environment. [ABSTRACT FROM AUTHOR]

The importance of the role of fibroblasts in cancer microenvironment is well-recognized. However, the relationship between fibroblasts and asbestos-induced lung cancer remains underexplored. To investigate the effect of the asbestos-related microenvironment on lung cancer progression, lung cancer cells (NCI-H358, Calu-3, and A549) were cultured in media derived from IMR-90 lung fibroblasts exposed to 50 mg/L asbestos (chrysotile, amosite, and crocidolite) for 24 h. The kinetics and migration of lung cancer cells in the presence of asbestos-exposed lung fibroblast media were monitored using a real-time cell analysis system. Proliferation and migration of A549 cells increased in the presence of media derived from asbestos-exposed lung fibroblasts than in the presence of media derived from normal lung fibroblasts. We observed no increase in proliferation and migration in lung cancer cells cultured in asbestos-exposed lung cancer cell medium. In contrast, increased proliferation and migration in lung cancer cells exposed to media from asbestos-exposed lung fibroblasts was observed for all types of asbestos. Media derived from lung fibroblasts exposed to other stressors, such as hydrogen peroxide and UV radiation didn’t show as similar effect as asbestos exposure. An enzyme-linked immunosorbent assay (ELISA)-based cytokine array identified interleukin (IL)-6 and IL-8, which show pleiotropic regulatory effects on lung cancer cells, to be specifically produced in higher amounts by the three types of asbestos-exposed lung fibroblasts than normal lung fibroblasts. Thus, the present study demonstrated that interaction of lung fibroblasts with asbestos may support the growth and metastasis of lung cancer cells and that chrysotile exposure can lead to lung cancer similar to that caused by amphibole asbestos (amosite and crocidolite). [ABSTRACT FROM AUTHOR]

The lentiviral Rev protein, which is a regulatory protein essential for virus replication, has been first studied in the human immunodeficiency virus type 1 (HIV-1). The main function of Rev is to mediate the nuclear exportation of viral RNAs. To fulfill its function, Rev shuttles between the cytoplasm and the nucleus. The Jembrana disease virus (JDV), a lentivirus, is the etiologic agent of the Jembrana disease which was first described in Bali cattle in Indonesia in 1964. Despite the high mortality rate associated with JDV, this virus remains poorly studied. Herein the subcellular distribution of JDV Rev, the nuclear and nucleolar localization signals (NLS and NoLS, respectively) and the nuclear export signal (NES) of the protein were examined. JDV Rev fused to the enhanced green fluorescent protein (EGFP) predominantly localized to the cytoplasm and nucleolus of transfected cells, as determined by fluorescence microscopy analyses. Through transfection of a series of deletion mutants of JDV Rev, it was possible to localize the NLS/NoLS region between amino acids (aa) 74 to 105. By substituting basic residues with alanine within this sequence, we demonstrated that the JDV Rev NLS encompasses aa 76 to 86, and is exclusively composed of arginine residues, whereas a bipartite NoLS was observed for the first time in any retroviral Rev/Rev-like proteins. Finally, a NES was identified downstream of the NLS/NoLS and encompasses aa 116 to 128 of the JDV Rev protein. The JDV Rev NES was found to be of the protein kinase A inhibitor (PKI) class instead of the HIV-1 Rev class. It also corresponds to the most optimal consensus sequence of PKI NES and, as such, is novel among lentiviral Rev NES. [ABSTRACT FROM AUTHOR]