Light Microscopy Images Research Articles

Label-Free Liquid Crystal Aptamer Sensors Based on Single-Stranded Nucleic Acid π-Structures for Detecting cTnI.

Cardiac troponin I (cTnI) is a highly sensitive and important serological marker for clinical diagnosis of myocardial injury. Its rapid detection is crucial for the early diagnosis of cardiovascular diseases such as acute myocardial infarction. In this study, based on nucleic acid molecular hybridization and aptamer-specific binding to target molecules, a label-free liquid crystal aptamer sensor based on single-stranded nucleic acid π-structures was developed and applied for the quantitative detection of cTnI. The CP1 and CP2 oligonucleotide chains, complementary to the bases at both ends of the aptamer, are covalently bonded to the sensor substrate via APTES and GA-mediated molecules. The aptamer forms a π-structure with CP1 and CP2 through nucleic acid hybridization, serving as a target molecule capture probe. When cTnI is present in the system, cTnI and the complementary oligonucleotide chains competitively bind with the aptamer, causing the breakdown of the π-structure within the sensor. This reinstates the long-range ordered alignment of the 5CB liquid crystal molecules within the sensor, enabling quantitative measurement of cTnI through variations in optical images. Experimental results show that within the range of 0.01 to 25 ng/mL for cTnI concentration, there is a linear correlation between the brightness area coverage (Br) in the polarized light microscopy images of the sensor and the logarithm of the cTnI concentration, with a correlation coefficient (r). The detection limit is 5.16 pg/mL. This method is label-free, simple to operate, and low-cost, with good specificity and a low detection limit, achieving cTnI detection in serum samples.

Application of convolutional neural networks and ensemble methods in the fiber volume content analysis of natural fiber composites

The incorporation of natural fibers into fiber-reinforced polymer composites (FRPC) has the potential to bolster their sustainability. A critical attribute of FRPC is the fiber volume content (FVC), a parameter that profoundly influences their thermo-mechanical characteristics. However, the determination of FVC in natural fiber composites (NFC) through manual analysis of light microscopy images is a labor-intensive process. In this work, it is demonstrated that the pixels from light microscopy images of NFC can be utilized to predict FVC using machine learning (ML) models. In this proof-of-concept investigation, it is shown that convolutional neural network-based models predict FVC with an accuracy required in polymer engineering applications, with a mean average error of 2.72% and an R2 coefficient of 0.85. Finally, it is shown that much simpler ML models, non-specialized in image recognition, besides being much easier and more efficient to optimize and train, can also deliver good accuracies required for FVC characterization, which not only contributes to the sustainability, but also facilitates the access of such models by researchers in regions with little computational resources. This study marks a substantial advancement in the area of automated characterization of NFC, and democratization of knowledge, offering a promising avenue for the enhancement of sustainable materials.

Thin-layer cervical sample evaluation: Conventional light microscopy versus digital whole-slide imaging.

Whole-slide imaging (WSI) has been adopted in many fields of pathology for education, quality assurance, and remote diagnostics. In 2021, the College of American Pathologists (CAP) updated guidelines to support pathology laboratories regarding the WSI systems validation process. However, the majority of published literature refers to histopathology rather than cytology. The aim of this study was to compare conventional light microscopy (CLM) and WSI in thin-layer cervical samples evaluation according to CAP guideline. A sample set of 64 thin-layer cervical specimens from women 25-64 years old who participated in cervical cancer screening programs in Tuscany was distributed among five cytologists at Institute for Cancer Research, Prevention and Clinical Network (Florence, Italy) for CLM analysis. After 2 weeks, the corresponding 64 digitally scanned slides were available at several magnifications for WSI evaluation. Substantial/near perfect agreement between CLM and WSI evaluation (0.77 ≥ κ ≥1) was observed for the negative for intraepithelial lesion or malignancy (NILM) class with concordance rates from 83.3% to 100%.Variability in concordance was observed among all the cytologists: 50%-85.7% for low-grade squamous intraepithelial lesion (LSIL), 47.1%-100% for high-grade squamous intraepithelial lesion (HSIL), 50%-100% for atypical glandular cells (AGCs) favors adenocarcinoma (ADK) with moderate/near perfect agreement (0.47 ≥ κ ≥1). Concordance and agreement rates were also variable within the "borderline" cytological categories of atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells cannot exclude an HSIL (ASC-H), and AGCs with lower or not computable kappa for some readers. The overall intralaboratory concordance between CLM and WSI was 92.9% with a near perfect agreement (κ =0.84) for NILM. Substantial agreement (κ ≥0.65) was assessed for LSIL, HSIL/squamous cell carcinoma, AGCs, and ADK categories whereas the agreement was lower (κ ≤0.39) for ASC-US and ASC-H. This study showed an overall substantial/near perfect agreement between CLM and WSI for all the cytological categories except the "borderline" ASC-US and ASC-H. Further progress in cytology WSI systems are needed before introducing it in routine diagnostics.

Analyzing the effectiveness of various coatings to mitigate photovoltaic modules soiling in desert climate

In this work, two different anti-soiling coatings (ASC) and anti-reflection coatings (ARC) were developed and tested under real operating conditions in Doha, Qatar, and compared with uncoated glass (UNC) to identify the best coating properties for mitigating solar photovoltaic (PV) module soiling. Soiling was measured in terms of loss in light transmittance, measured by UV–Vis spectroscopy, and increase in surface coverage area, measured using optical microscopy. The results indicate a decrease in light transmittance and an increase in surface coverage area over time due to dust accumulation. ASC coupons show higher hydrophilicity and, therefore, less soiling. ASC/ARC leads to a higher loss in light transmittance, while UNC results in a higher surface coverage area compared to the other coupons. Over 25 weeks of outdoor exposure and 12 cleaning cycles, analysis of 13,320 light microscopy images revealed mean dust surface coverage areas for ASC, ASC/ARC, and UNC glass coupons of 14.6 %, 13.6 %, and 15.8 %, respectively. Unlike the ASC/ARC and UNC coupons, the ASC recovers to its initial condition immediately after cleaning. Therefore, ASC was found to maintain a cleaner glass surface compared to ASC/ARC and UNC. However, the differences in transmittance loss among the three coupons are not significant. The results were compared to findings from similar studies conducted in Saudi Arabia to evaluate regional differences in coatings performance. For the same coatings, a significant light transmittance loss was measured in Doha (Qatar) compared to Uyaynah (Saudi Arabia). The combination of high humidity and heavy dust in Qatar's extreme conditions reduces the effectiveness of both ASC/ARC and ASC coatings.

RVE Simulations of short fiber reinforced polyamide: Direct and inverse matrix parameter identification in view of the semi-crystalline polymer structure

This paper investigates the modeling capabilities of computational homogenization for the mechanical behavior of short fiber reinforced polyamide. Simulations on a representative volume element (RVE) with elastic fibers and an elastic–plastic matrix are compared to tensile experiments on specimens taken in parallel and transversal direction from injection molded plates. In view of the semi-crystalline polymer structure, focus is put on identifying the matrix parameters through two alternative methods:First, the matrix parameters are identified directly using tensile experiments on a non-nucleated and a nucleated unreinforced polyamide. In the RVE computations based on the non-nucleated grade, the composite stress–strain behavior is somewhat underestimated, and with the nucleated grade, the behavior is slightly overestimated. To explain this, the semi-crystalline polymer structure is studied. Polarized light microscopy images reveal that the non-nucleated grade has a coarser and the nucleated grade a finer spherulite structure, compared to the matrix present in the composite. However, the degree of crystallinity measured by differential scanning calorimetry is in a similar range.As an alternative, the matrix parameters are identified inversely by fitting the RVE model to the composite tensile experiments. As uncertainties with respect to the matrix material as well as possibly remaining simplifications in the micromechanical model are compensated for, this reverse engineering approach allows for a very good fit.

Clearing-enabled light sheet microscopy as a novel method for three-dimensional mapping of the sensory innervation of the mouse knee.

A major barrier that hampers our understanding of the precise anatomic distribution of pain sensing nerves in and around the joint is the limited view obtained from traditional two dimensional (D) histological approaches. Therefore, our objective was to develop a workflow that allows examination of the innervation of the intact mouse knee joint in 3D by employing clearing-enabled light sheet microscopy. We first surveyed existing clearing protocols (SUMIC, PEGASOS, and DISCO) to determine their ability to clear the whole mouse knee joint, and discovered that a DISCO protocol provided the optimal transparency for light sheet microscopy imaging. We then modified the DISCO protocol to enhance binding and penetration of antibodies used for labeling nerves. Using the pan-neuronal PGP9.5 antibody, our protocol allowed 3D visualization of innervation in and around the mouse knee joint. We then implemented the workflow in mice intra-articularly injected with nerve growth factor (NGF) to determine whether changes in the nerve density can be observed. Both 3D and 2D analytical approaches of the light sheet microscopy images demonstrated quantifiable changes in midjoint nerve density following 4 weeks of NGF injection in the medial but not in the lateral joint compartment. We provide, for the first time, a comprehensive workflow that allows detailed and quantifiable examination of mouse knee joint innervation in 3D.

Employing Synchrotron X-ray Scattering and Microscopy to Explore Microstructural Mysteries in Bioresorbable Vascular Scaffolds

Crystal structure and morphology dictate the mechanical, thermal, and degradation properties of poly l-lactide (PLLA), the structural polymer of the first clinically approved bioresorbable vascular scaffolds (BVS). New experimental methods are developed to reveal the underlying mechanisms governing structure formation during the crimping step of the BVS manufacturing process. Our research specifically examines the “U-bends” – the region where the curvature is highest and stress is maximised during crimping, which can potentially lead to failure of the device with dramatic consequences on patient life. A custom-made crimping rig operated at a synchrotron beamline enabled collection of wide- and small-angle X-ray scattering (WAXS/SAXS) to probe local variations of the polymer morphology as a function of position in the crest of multiple U-bends with 5 μm resolution in situ after crimping and expansion. Additionally, polarised light microscopy (PLM) images of these deformed U-bends revealed areas with varying stress distribution developed during crimping and expansion. These variations were dependant on the initial biaxial stretching processing step. The integrated X-ray scattering-microscopy approach offered a comprehensive work-flow for uncovering the intricate relationship between processing conditions and the corresponding spatially-resolved semicrystalline morphology of a BVS. Statement of SignificanceThis research introduces a new method for gaining critical insights into the structural changes that occur during the manufacturing process of bioresorbable vascular scaffolds (BVS). The crimping and expansion of poly l-lactide (PLLA) – the structural material of BVS – are sequential manufacturing steps characterised by highly non-linear deformations at temperature conditions that remain unexplored.By utilising synchrotron X-ray scattering techniques alongside polarised light microscopy, we have developed new experimental methods to uncover the mechanisms governing structure formation during processing. This innovative approach not only deepens our understanding of the relationship between processing conditions and polymer morphology but also establishes the foundation for real-time observation methods during crimping and expansion. By improving the design and performance of BVS, this study has the potential to advance cardiovascular treatments and improve patient safety, making it highly relevant and impactful to both scientific research and clinical applications.

Microstructure evolution of AlSi10Mg alloy in RAP process

Abstract With its computer-aided layer-by-layer production approach, additive manufacturing (AM) and powder bed laser fusion (PBLF) paved the way to produce metallic parts more precisely than any other manufacturing technique. However, the combinability and the interaction of this relatively new manufacturing technique with the other near-net shape production techniques is still a mystery. In this study, the recrystallization and partial melting (RAP) behavior, which is a feedstock production approach for semisolid forming methods, investigated on AlSi10Mg parts produced by PBLF and conventional casting were compared in terms of microstructural and hardness evaluations. After the reheating process, the globalization of Si particles and the breakdown of the Si network around the melt pools were displayed with light microscope and scanning electron microscope (SEM) images. The hardness values of the PBLF as-fabricated specimens were found to be significantly higher than the as-cast specimens; however, the values were almost equaled after the RAP treatment and even got lower on the bottom and top regions of the PBLF samples after 20 min of reheating because of the enlarged shrinkage porosities and the coarsened morphology.

An ultrastructure analysis of the developing human anterior cruciate ligament tibial enthesis.

This study aimed to investigate the ultrastructural anatomy of the developing ACL tibial enthesis. We hypothesized that enthesis architecture would progressively mature and remodel, eventually resembling that of the adult by the early postnatal stage. Five fresh-frozen human pediatric cadaveric knees aged 1-36 months underwent anatomical dissection to harvest the ACL insertion and underlying tibial chondroepiphysis. The samples were prepared for scanning electron microscopy (SEM) to examine the ultrastructural anatomy of the enthesis and underwent histological staining for circular polarized light (CPL) and light microscopy imaging. SEM analysis of the 1- and 8-month-old samples revealed a shallow interdigitation between the dense fibrous (ligamentous) tissue and unmineralized chondrogenic tissues, with a minimal transition zone. By 11-month, a more complex transition zone was present. By age 19- and 36-month-old, a progressively more complex and defined fibrocartilage zone was observed. CPL analysis revealed distinct collagen fiber continuity, alignment, and organization changes over time. By 19 and 36 months, the samples exhibited complex fiber arrangements and a progression toward uniform fiber orientation. Similarly, histological analysis demonstrated progressive remodeling of the enthesis with increasing age. Our results suggest that the ACL enthesis of the developing knee begins to mimic that of an adult as early as 19 months of age, as a more complex transition between ligamentous and chondro-epiphyseal tissue can be appreciated. We hypothesize that the observed changes are likely due to mechanical loading of the enthesis with the onset of weightbearing. Future investigations of ACL reconstruction and repair will benefit from improved understanding of the chondro-epiphyseal/ACL regions.

Halamphorahampyeongensis sp. nov. (Amphipleuraceae, Bacillariophyceae), a new marine benthic diatom from a tidal mudflat in Hampyeong Bay, South Korea.

The abundance and variety of benthic diatoms inhabiting tidal flats is widely acknowledged, although it has received relatively less attention than other research areas. In this investigation, we provide a formal description of a benthic diatom found in the tidal mudflat of South Korea, based on morphological and molecular characteristics and the similarities and differences between Halamphorahampyeongensis sp. nov., with morphologically similar Halamphora species are also discussed. Morphological characteristics are described from light and electron microscopy images. H.hampyeongensis is distinguished by its wide ventral sides of the valve, small and rounded areolae present across the whole valve face, and dense dorsal striae biseriate (34-38 in 10 μm). Phylogenetic analysis based on 18S rDNA and rbcL sequence data revealed that H.hampyeongensis is related to H.montana, H.mosensis, and H.specensa. The results (morphometric and molecular) provide sufficient elements to support and propose this taxon as a new species.

Correlative microscopy - illuminating the endomembrane system of plant seeds.

The endomembrane system of cereal seed endosperm is a highly plastic and dynamic system reflecting the high degree of specialization of this tissue. It is capable of coping with high levels of storage protein synthesis and undergoes rapid changes to accommodate these storage proteins in newly formed storage organelles such as endoplasmic reticulum-derived protein bodies or protein storage vacuoles. The study of endomembrane morphology in cereal endosperm is challenging due to the amount of starch that cereal seeds accumulate and the progressive desiccation of the tissue. Here, we present a comprehensive study of the endomembrane system of developing barley endosperm cells, complemented by correlative light and electron microscopy (CLEM) imaging. The use of genetically fused fluorescent protein tags in combination with the high resolution of electron microscopy brings ultrastructural research to a new level and can be used to generate novel insights in cell biology in general and in cereal seed research in particular.

Concordance of Whole-Slide Imaging and Conventional Light Microscopy for Assessment of Pathologic Response Following Neoadjuvant Therapy for Lung Cancer

Pathologic response is an end point in many ongoing clinical trials for neoadjuvant regimens, including immune checkpoint blockade and chemotherapy. Whole-slide scanning of glass slides generates high-resolution digital images and allows for remote review and potential measurement with image analysis tools, but concordance of pathologic response assessment on digital scans compared with that on glass slides has yet to be evaluated. Such a validation goes beyond previous concordance studies, which focused on establishing surgical pathology diagnoses, as it requires quantitative assessment of tumor, necrosis, and regression. Further, as pathologic response assessment is being used as an end point, such concordance studies have regulatory implications. The purpose of this study was 2-fold, which was as follows: first, to determine the concordance between pathologic response assessed on glass slides and that assessed on digital scans, and second, to determine if pathologists benefited from using measurement tools when determining pathologic response. To that end, hematoxylin and eosin–stained glass slides from 64 non–small cell lung carcinoma specimens were visually assessed for percent residual viable tumor (%RVT). The sensitivity and specificity for digital vs glass reads of pathologic complete response (0% RVT) and major pathologic response (≤10% RVT) were all >95%. When %RVT was considered as a continuous variable, the intraclass correlation coefficient of digital vs glass reads was 0.94. The visual assessments of pathologic response were supported by pathologist annotations of residual tumor and tumor bed areas. In a separate subset of hematoxylin and eosin–stained glass slides, several measurement approaches to quantifying %RVT were performed. Pathologist estimates strongly reflected measured %RVT. This study demonstrates the high level of concordance between glass slides evaluated using light microscopy and digital whole-slide images for pathologic response assessments. Pathologists did not require measurement tools to generate robust %RVT values from slide annotations. These findings have broad implications for improving clinical workflows and multisite clinical trials.

Studies on type material from Kützing’s diatom collection VIII. Species assigned to the genera Epithemia Brébisson ex Kützing and Rhopalodia O. Müller

We detail the diatom taxa described by F.T. Kützing across three of his publications that have been referred to the diatom genera Epithemia and Rhopalodia. Taxa considered include Frustulia adnata, F. picta, E. alpestris, E. musculus, E. porcellus, E. proboscidea, E. sorex, E. ventricosa, E. vertagus, E. ventricosa β. gregaria and E. reticulata. For these 12 taxa, this represents the first time Kützing’s specimens have been illustrated with light microscopy. Most of the material consulted here is from the Natural History Museum, London, though we also reviewed material from the Academy of Natural Sciences for E. adnata. We present light microscope images of the Kützing taxa and complement these observations with images of the type slides, material packets and both herbarium (unpublished) and published drawings. In some cases, modern concepts of Kützing species are supported, but in others it is likely that species concepts will be changed by the specimens in the type material. Lectotypes are designated for Epithemia adnata and Epithemia reticulata.

Reevaluation of pollen differentiation in Altingiaceae: Challenges in distinguishing deciduous (Liquidambar) and evergreen (Altingia) types using multivariate statistics and machine learning

Altingiaceae, a family of woody plants, comprising evergreen Altingia and deciduous Liquidambar groups, exhibits distinct leaf morphology, yet both groups overlap in geographical range and climatic conditions. While some tropical Altingia species are confined to Myanmar, Thailand, Cambodia and India without Liquidambar, and some temperate Liquidambar species to northern China without Altingia. Their fossil pollen have significant implications in reconstructing palaeoclimate and historical biogeography, based on classification of Altingia-type and Liquidambar-type. However, the results of previous studies to differentiate pollen types of evergreen Altingia and deciduous Liquidambar were based on limited pollen specimens. Therefore pollen morphology of Altingiaceae and differentiation of above mentioned types needs reevaluation using more specimens from wider geographical range.In this study, we present new findings on Altingiaceae pollen morphology from extensive collection of specimens and reassess the diagnostic features to distinguish evergreen and deciduous types. To improve the credibility of palaeoecological and palaeoclimatic interpretations, we applied multivariate statistical analyses to pollen size, number of pores, pollen wall thickness, and size and density of ornamental elements from light microscopy (LM) and Scanning Electronic Microscope (SEM) images. Additionally, random forest classification models were applied to test the accuracy of differentiating Altingiaceae pollen types. Our results reveal significant morphological overlap between the pollen of evergreen Altingia and deciduous Liquidambar, with classification models showing limited accuracy and explainability. Thus, fossil pollen of Altingiaceae cannot be confidently classified into evergreen or deciduous types, highlighting challenges in using their pollen morphology for taxonomic classification in palaeoecological research.

Foliar epidermal micro-morphology of selected brinjal varieties: A study on leaf surface characteristics

Studies on the leaf epidermal morphological characteristics were conducted on the three varieties of brinjal (Solanum melongena L.) belonging to the family Solanaceae. The varieties used in the study were Bhavani Gold, Purple Round and Haritha. Fresh mature brinjal leaves were randomly selected from the three experimental genotypes, and the foliar epidermal features were studied by using light microscopy and SEM imaging. Measurements were taken with an ocular and stage micrometer using 10X and 40X magnifications. The photographs were taken from mounted slides. The stomatal count and each measurement represent the average of ten readings. The upper and lower epidermal surfaces of leaf were studied for stomatal features including, stomatal distribution, stomatal type, stomatal index, stomatal pore length, stomatal pore width, guard cell length, guard cell width, subsidiary cell length and width, epidermal cell type, epidermal cell length and width, trichome type, trichome length and width. Amphistomatous, anisocytic, and with more stomata on the lower epidermal surface than the upper epidermal surface were present in all leaf varieties. On the leaves, the abaxial stomatal index ranged from 68.19 mm-2 (Bhavani Gold) to 56.67 mm-2 (Purple Round) to 58.8 mm-2 (Haritha). While on the adaxial surface, it varied from 59.67 mm-2 (Bhavani Gold), 53.46 mm-2 (Purple Round) and 56.67 mm-2 (Haritha). The three varieties of this study exhibited similarities in the pattern of epidermal cell on the adaxial and abaxial surfaces and were irregular in shape. Among the three varieties epidermal cell become irregular in outline and trichomes were present on both leaf surfaces. It becomes stellate, non-glandular, glandular and 8-12 in number. The highest trichome length was obtained in Bhavani Gold (63 μm) followed by Haritha (57.5 μm) and Purple Round (43 μm) on the upper epidermis. On the lower epidermal region maximum length was obtained in Bhavani Gold (65.5 μm) followed by Haritha (53 μm) and Purple Round (44 μm).

A Pharmacognostic, Phytochemical Study of Godhuma Bija (Triticum Aestivum Linn.)

Background: Godhuma Bija (Triticum aestivum Linn.)are used for various conditions of ailments in traditional systems of medicine since ancient times. Pharmacognostic and phytochemical studies make the drugs more standardized and evidence based. Aims: This study was designed to lay down the various pharmacognostic and phytochemical standards which will be helpful to ensure the purity, safety, and efficacy of this medicinal plant. Materials and Methods: Various methods including macroscopic, microscopic, physicochemical and phytochemical methods were applied to determine the diagnostic features for the identification and standardization of intact and powdered drug of Godhuma Bija (Triticum aestivum Linn.) Results: The shape, colour, odour and surface characteristics were determined for the intact drug and powdered materials of Godhuma Bija (Triticum aestivum Linn.) Light and electron microscope images of cross-section of stamen and powdered microscopy revealed useful diagnostic features. Phytochemical, physicochemical analysis of powdered drug proved useful to differentiate the powdered drug material. High performance thin layer chromatography analysis showed the presence of important phytoconstituents. Conclusion: Morphology as well as various pharmacognostic aspects of Godhuma Bija (Triticum aestivum Linn.) of the plant were studied and have been described here along with phytochemical and physicochemical studies, which will help in authentication and quality control.

Large-volume fully automated cell reconstruction generates a cell atlas of plant tissues.

The geometric shape and arrangement of individual cells play a role in shaping organ functions. However, analyzing multicellular features and exploring their connectomes in centimeter-scale plant organs remain challenging. Here, we established a set of frameworks named Large-Volume Fully Automated Cell Reconstruction (LVACR), enabling the exploration of three-dimensional (3D) cytological features and cellular connectivity in plant tissues. Through benchmark testing, our framework demonstrated superior efficiency in cell segmentation and aggregation, successfully addressing the inherent challenges posed by light sheet fluorescence microscopy (LSFM) imaging. Using LVACR, we successfully established a cell atlas of different plant tissues. Cellular morphology analysis revealed differences of cell clusters and shapes in between different poplar (P. simonii Carr. and P. canadensis Moench.) seeds, whereas topological analysis revealed that they maintained conserved cellular connectivity. Furthermore, LVACR spatiotemporally demonstrated an initial burst of cell proliferation, accompanied by morphological transformations at an early stage in developing the shoot apical meristem. During subsequent development, cell differentiation produced anisotropic features, thereby resulting in various cell shapes. Overall, our findings provided valuable insights into the precise spatial arrangement and cellular behavior of multicellular organisms, thus enhancing our understanding of the complex processes underlying plant growth and differentiation.

Overview of the Caucasian Perla Geoffroy, 1762 (Plecoptera: Perlidae) based on morphological and molecular data with description of two new species.

Six species of Caucasian Perla are reviewed, and diagnostic morphological characteristics of all stages of development (where possible) are described, supplemented, and illustrated in detail with comparative light microscope and scanning electron microscopy images. The DNA barcoding of five species is presented. Two new morphologically and genetically distinct species, Perla schapsugica sp. nov. and Perla palatovi sp. nov., are described for both sexes and all life stages in the North Caucasus, Russia, Krasnodar Kray. Reinstatement of Perla persica Zwick, 1975, as a valid species distinct from P. caucasica Guérin-Méneville, 1843, is proposed. A new record of P. persica is reported for the Greater Caucasus, Russia, North-Ossetia-Alania for the first time. Morphologically, these two latter species can be separated in male adults by the shape of the hemitergal hook on terga X, an additional ventral brush on the penis of P. caucasica, wing length, and color.

SynBot is an open-source image analysis software for automated quantification of synapses

The formation of precise numbers of neuronal connections, known as synapses, is crucial for brain function. Therefore, synaptogenesis mechanisms have been one of the main focuses of neuroscience. Immunohistochemistry is a common tool for visualizing synapses. Thus, quantifying the numbers of synapses from light microscopy images enables screening the impacts of experimental manipulations on synapse development. Despite its utility, this approach is paired with low-throughput analysis methods that are challenging to learn, and the results are variable between experimenters, especially when analyzing noisy images of brain tissue. We developed an open-source ImageJ-based software, SynBot, to address these technical bottlenecks by automating the analysis. SynBot incorporates the advanced algorithms ilastik and SynQuant for accurate thresholding for synaptic puncta identification, and the code can easily be modified by users. The use of this software will allow for rapid and reproducible screening of synaptic phenotypes in healthy and diseased nervous systems.

Biological control of ultra-skeleton mineralization in coral

Abstract Understanding the mineralization of coral is significant for the formation of coral reefs and paleoclimatic reconstructions. However, the fundamental mechanisms involved in biomineralization are poorly understood. A combination of Raman spectral and cross-polarized reflected light microscopy imaging was used to examine the three-dimensional spatial distribution of the skeletal ultrastructures and their associated mineral, organic, and water chemistry in coral, which enable insight into the spatial growth features of the ultrastructures and possible formation processes. A possible mechanism is proposed that controls the formation of skeletal ultrastructures, which likely involves compartmentalized calcifying cells and their related cellular activities. This could clarify the association between coral skeletal mineralization and biology, and it may be beneficial to better protection and application of coral reefs.