Quantitative Morphological Analysis Research Articles

Image-Based Quantitative Analysis of Epidermal Morphology in Wild Potato Leaves.

The epidermal leaf patterns of plants exhibit remarkable diversity in cell shapes, sizes, and arrangements, driven by environmental interactions that lead to significant adaptive changes even among closely related species. The Solanaceae family, known for its high diversity of adaptive epidermal structures, has traditionally been studied using qualitative phenotypic descriptions. To advance this, we developed a workflow combining multi-scale computer vision, image processing, and data analysis to extract digital descriptors for leaf epidermal cell morphology. Applied to nine wild potato species, this workflow quantified key morphological parameters, identifying descriptors for trichomes, stomata, and pavement cells, and revealing interdependencies among these traits. Principal component analysis (PCA) highlighted two main axes, accounting for 45% and 21% of variance, corresponding to features such as guard cell shape, trichome length, stomatal density, and trichome density. These axes aligned well with the historical and geographical origins of the species, separating southern from Central American species, and forming distinct clusters for monophyletic groups. This workflow thus establishes a quantitative foundation for investigating leaf epidermal cell morphology within phylogenetic and geographic contexts.

Osteosarcoma stem cells resist chemotherapy by maintaining mitochondrial dynamic stability via DRP1.

Osteosarcoma malignancy exhibits significant heterogeneity, comprising both osteosarcoma stem cells (OSCs) and non‑OSCs. OSCs demonstrate increased resistance to chemotherapy due to their distinctive cellular and molecular characteristics. Alterations in mitochondrial morphology and homeostasis may enhance chemoresistance by modulating metabolic and regulatory processes. However, the relationship between mitochondrial homeostasis and chemoresistance in OSCs remains to be elucidated. The present study employed high‑resolution microscopy to perform multi‑layered image reconstructions for a quantitative analysis of mitochondrial morphology. The results indicated that OSCs exhibited larger mitochondria in comparison with non‑OSCs. Furthermore, treatment of OSCs with cisplatin (CIS) or doxorubicin (DOX) resulted in preserved mitochondrial morphological stability, which was not observed in non‑OSCs. This finding suggested a potential association between mitochondrial homeostasis and chemoresistance. Further analysis indicated that dynamin‑related protein 1 (DRP1) might play a pivotal role in maintaining the stability of mitochondrial homeostasis in OSCs. Depletion of DRP1 resulted in the disruption of mitochondrial stability when OSCs were treated with CIS or DOX. Additionally, knocking out DRP1 in OSCs led to a reduction in chemoresistance. These findings unveil a novel mechanism underlying chemoresistance in osteosarcoma and suggest that targeting DRP1 could be a promising therapeutic strategy to overcome chemoresistance in OSCs. This provided valuable insights for enhancing treatment outcomes among patients with osteosarcoma.

Plant cell wall enzymatic deconstruction: Bridging the gap between micro and nano scales

Understanding lignocellulosic biomass resistance to enzymatic deconstruction is crucial for its sustainable conversion into bioproducts. Despite scientific advances, quantitative morphological analysis of plant deconstruction at cell and tissue scales remains under-explored. In this study, an original pipeline is devised, involving four-dimensional (space + time) fluorescence confocal imaging, and a novel computational tool, to track and quantify deconstruction at cell and tissue scales. By applying this pipeline to poplar wood, dynamics of cellular parameters was computed and cellulose conversion during enzymatic deconstruction was measured. Results showed that enzymatic deconstruction predominantly impacts cell wall volume rather than surface area. Additionally, a negative correlation was observed between pre-hydrolysis compactness measures and volumetric cell wall deconstruction rate, whose strength was modulated by enzymatic activity. Results also revealed a strong positive correlation between average volumetric cell wall deconstruction rate and cellulose conversion rate. These findings link key deconstruction parameters across nano and micro scales.

3D morphometry of endothelial cells angiogenesis in an extracellular matrix composite hydrogel

Human umbilical vein endothelial cells (HUVECs) play a fundamental role in angiogenesis. Herein, we introduce digital holographic microscopy (DHM) for the 3D quantitative morphological analysis of HUVECs in extracellular matrix (ECM)-based biomaterials as an angiogenesis model. The combination of volumetric information from DHM and the physicochemical and cytobiocompatibility data provided by fluorescence microscopy and cytology offers a comprehensive understanding of the angiogenesis-related parameters of HUVECs within the ECM. DHM enables label-free, non-contact, and non-invasive 3D monitoring of living samples in real time, in a quantitative manner. In this study, the human amniotic membrane (HAM) is decellularized, pulverized, and combined with sodium alginate hydrogel to provide an in vitro substrate for modeling HUVEC angiogenesis. Our results demonstrate that modifying alginate hydrogel with HAM enhances its biofunctionality due to the presence of ECM components. Moreover, the DHM results reveal an increase in its porous properties, which, in turn, aids in interpreting the tubulation results.

Experimental study on the influence of particle morphology on compression characteristics of coral sand in South China Sea

Particle morphology is an important physical parameter that influences the engineering properties of coral sand, especially its compressibility. According to the quantitative analysis of particle morphology via the dynamic image analysis technology, a series of one-dimensional compression tests on coral sand and quartz sand with different particle morphology and densities were performed to reveal the effect of particle morphology on the compressibility of coral sand. The results demonstrated that the morphology of particles can be quantified using the shape-angularity group indicator (SAGI), whereas the SAGI of coral sand was greater than that of quartz sand. Coral sand exhibited a yield stress ranging from 1 to 2.5 MPa, and compressive deformations in both coral sand and quartz sand were dominated by irreversible plastic deformation. As the SAGI and densification increased, the number of particle coordination increased while the contact stress between grains decreased, which caused the yield stress of coral sand specimens to increase as well as the compression index and particle crushing rate to decrease. The SAGI of quartz sand particles remained relatively unchanged while that of coral sand particles gradually decreased after crushing, i.e., the coral sand particles developed a more regular shape after crushing.

Characterization of Photo-Crosslinked Methacrylated Type I Collagen as a Platform to Investigate the Lymphatic Endothelial Cell Response

Despite chronic fibrosis occurring in many pathological conditions, few in vitro studies examine how fibrosis impacts lymphatic endothelial cell (LEC) behavior. This study examined stiffening profiles of PhotoCol®—commercially available methacrylated type I collagen—photo-crosslinked with the photoinitiators: Lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), Irgacure 2959 (IRG), and Ruthenium/Sodium Persulfate (Ru/SPS) prior to evaluating PhotoCol® permeability and LEC response to PhotoCol® at stiffnesses representing normal and fibrotic tissues. Ru/SPS produced the highest stiffness (~6 kilopascal (kPa)) for photo-crosslinked PhotoCol®, but stiffness did not change with burst light exposures (30 and 90 s). The collagen fibril area fraction increased, and dextran permeability (40 kilodalton (kDa)) decreased with photo-crosslinking, showing the impact of photo-crosslinking on microstructure and molecular transport. Human dermal LECs on softer, uncrosslinked PhotoCol® (~0.5 kPa) appeared smaller with less prominent vascular endothelial (VE)-cadherin (cell–cell junction) expression compared to LECs on stiffer PhotoCol® (~6 kPa), which had increased cell size, border irregularity, and VE-cadherin thickness (junction zippering) that is consistent with LEC morphology in fibrotic tissues. Our quantitative morphological analysis demonstrates our ability to produce LECs with a fibrotic phenotype, and the overall study shows that PhotoCol® with Ru/SPS provides the necessary physical properties to systematically study LEC responses related to capillary growth and function under fibrotic conditions.

3D X-ray Tomography Analysis of Mg–Si–Zn Alloys for Biomedical Applications: Elucidating the Morphology of the MgZn Phase

Temporary metal implants, made from materials like titanium (Ti) or stainless steel, can cause metabolic issues, raise toxicity levels within the body, and negatively impact the patient’s long-term health. This necessitates a subsequent operation to extract these implants once the healing process is complete or when they are outgrown by the patient. In contrast, medical devices fabricated from absorbable alloys have the advantage of being biodegradable, allowing them to be naturally absorbed by the body once they have fulfilled their role in facilitating tissue healing. Among the various absorbable alloy systems studied, magnesium (Mg) alloys stand out due to their biocompatibility, mechanical properties, and corrosion behavior. The existing literature on absorbable Mg alloys highlights the effectiveness of silicon (Si) and zinc (Zn) additions in improving mechanical properties and controlling corrosion susceptibility; however, there is a lack of comprehensive quantitative morphological analysis of the intermetallic phases within these alloy systems. The quantification of the complex morphology of intermetallic particles is a challenging task and has significant implications for the micromechanical properties of the alloys. This study, therefore, aims to introduce a robust set of morphometric parameters for evaluating the morphology of intermetallic phases within two as-cast Mg alloys with Si and Zn additions. X-ray Computed Tomography (XCT) was used to capture the 3D tomographic data of the alloys, and a novel pair of morphological parameters (ratio of convex hull to particle volume and convex hull sphericity) was applied to the 3D tomographic data to assess the MgZn phase formed in the two alloys. In addition to the impact of composition, the effect of solidification rate on the morphological parameters was also studied. Furthermore, Scanning Electron Microscopy (SEM) and Energy-Dispersive Spectroscopy (EDS) were employed to gather detailed 2D microstructural and compositional information on the intermetallics. The comprehensive characterization reveals that the morphological complexity and size distribution of the MgZn phase are influenced by both compositional changes and the solidification rate. However, the change in MgZn intermetallic particle morphology with size was found to follow a predictable trend, which was relatively agnostic of the chosen casting conditions.

Study on thermal crack characteristics of granite in Shandong Province, China under different temperatures and heating/cooling treatments

The qualitative and quantitative analysis of crack initiation characteristics, extended procedure, and crack network morphology of rock under different temperatures and heating/cooling paths was conducted in order to delve deeper into the mechanism of thermal cracking, and the analytical perspectives mainly include microstructure, thermodynamics, and mineral crystal properties. Research results indicate that 200–300 °C and 600 °C are two obvious mutation points of water-cooled samples, while air-cooled samples show mutation points around 600 °C. The initiation positions of thermal cracking under all temperature paths are relatively similar, with intergranular crack located between particles at the edge of the sample or intercrystalline cracks in the middle of feldspar or quartz aggregates. Slow heating and rapid cooling contribute to the complexity of the thermal crack network, and rapid heating can form larger sized thermal cracks. The process of thermal cracking can be divided into three stages: initiation of small cracks, joint development of main cracks and small cracks, and interconnection of cracks to form a network structure. The thermal cracking mechanism is attributed to differences in expansion and thermal conductivity among various crystals.

Quantification and analysis study for the morphological characteristics of urban waterbodies: Evidences from 42 cases in China

Influenced by the water design principle of Chinese classical garden, the design of urban waterbodies (lakes, ponds and wetlands) in China typically exhibits natural-like morphological characteristics, but what similarities and differences among these characteristics remain unclear. Also, the morphology of urban waterbodies is mostly designed with beauty and lacks quantitative research and methodological support, which is not favourable to the exploration of the correlation between waterbody morphology and ecology. In this study, 11 quantifiable morphological indicators are selected from water surface, shoreline complexity and water-land spatial morphology. A Grasshopper-based model for waterbody morphology quantification and analysis was constructed and applied to 42 cases in China. Combined with frequency, correlation and PCA analysis, the cases and indicators with higher composite scores generally showed more natural water surface morphology, island distribution, and diversified land and water spaces. The thresholds for the more representative indicators are 0.10–0.60 for the circularity ratio (C), 1.01–1.15 for the shoreline fractal dimension (D0), 1.58–29.42 for the island nearest-neighbour index (NN), 1.20–2.50 for the shoreline development index (SDI), 0.20–0.55 for the compactness ratio (Rc) and 0.30–0.85 for the unit water surface morphological difference ratio (ΔB). Focusing on these indicators, along with ΔB and I which describe spatial inclusiveness and island distribution, their regression equations and threshold ranges provide a primary basis for developing waterbody morphology designs and generative algorithms. This methodology has been experimentally applied in the design of a case waterbody in Suqian City Park. Overall, this study constructs and validates the feasibility of a Grasshopper-based algorithm for the quantitative analysis and morphogenetic design of waterbody morphology. It demonstrates the potential of this method for understanding complex waterbody morphology and water-land spaces, and supports a new digital methodology for the optimisation of waterscape design.

The design of a sample rapid magnetic resonance imaging (MRI) acquisition protocol supporting assessment of multiple articular tissues and pathologies in knee osteoarthritis

ObjectiveThis expert opinion paper proposes a design for a state-of-the-art magnetic resonance image (MRI) acquisition protocol for knee osteoarthritis clinical trials in early and advanced disease. Semi-quantitative and quantitative imaging endpoints are supported, partly amendable to automated analysis. Several (peri-) articular tissues and pathologies are covered, including synovitis. MethodA PubMed literature search was conducted, with focus on the past 5 years. Further, osteoarthritis imaging experts provided input. Specific MRI sequences, orientations, spatial resolutions and parameter settings were identified to align with study goals. We strived for implementation on standard clinical scanner hardware, with a net acquisition time ≤30 ​min. ResultsShort- and long-term longitudinal MRIs should be obtained at ≥1.5T, if possible without hardware changes during the study. We suggest a series of gradient- and spin-echo-sequences, supporting MOAKS, quantitative analysis of cartilage morphology and T2, and non-contrast-enhanced depiction of synovitis. These sequences should be properly aligned and positioned using localizer images. One of the sequences may be repeated in each participant (re-test), optimally at baseline and follow-up, to estimate within-study precision. All images should be checked for quality and protocol-adherence as soon as possible after acquisition. Alternative approaches are suggested that expand on the structural endpoints presented. ConclusionsWe aim to bridge the gap between technical MRI acquisition guides and the wealth of imaging literature, proposing a balance between image acquisition efficiency (time), safety, and technical/methodological diversity. This approach may entertain scientific innovation on tissue structure and composition assessment in clinical trials on disease modification of knee osteoarthritis.

The morphological discrepancy of neuromuscular junctions between bilateral paraspinal muscles in patients with adolescent idiopathic scoliosis: A quantitative immunofluorescence assay.

Prior studies suggested that neuromuscular factors might be involved in the pathogenesis of adolescent idiopathic scoliosis (AIS). The neuromuscular junction (NMJ) is the important pivot where the nervous system interacts with muscle fibers, but it has not been well characterized in the paraspinal muscles of AIS. This study aims to perform the quantitative morphological analysis of NMJs from paraspinal muscles of AIS. AIS patients who received surgery in our center were prospectively enrolled. Meanwhile, age-matched congenital scoliosis (CS) and non-scoliosis patients were also included as controls. Fresh samples of paraspinal muscles were harvested intraoperatively. NMJs were immunolabeled using different antibodies to reveal pre-synaptic neuronal architecture and post-synaptic motor endplates. A confocal microscope was used to acquire z-stack projections of NMJs images. Then, NMJs images were analyzed on maximum intensity projections using ImageJ software. The morphology of NMJs was quantitatively measured by a standardized 'NMJ-morph' workflow. A total of 21 variables were measured and compared between different groups. A total of 15 AIS patients, 10 CS patients and 5 normal controls were enrolled initially. For AIS group, NMJs in the convex side of paraspinal muscles demonstrated obviously decreased overlap when compared with the concave side (34.27% ± 8.09% vs. 48.11% ± 10.31%, p = 0.0036). However, no variables showed statistical difference between both sides of paraspinal muscles in CS patients. In contrast with non-scoliosis controls, both sides of paraspinal muscles in AIS patients demonstrated significantly smaller muscle bundle diameters. This study first elucidated the morphological features of NMJs from paraspinal muscles of AIS patients. The NMJs in the convex side showed smaller overlap for AIS patients, but no difference was found in CS. This proved further evidence that neuromuscular factors might contribute to the mechanisms of AIS and could be considered as a novel potential therapeutic target for the treatment of progressive AIS.

AI-assisted deep learning segmentation and quantitative analysis of X-ray microtomography data from biomass ashes

X-ray microtomography is a non-destructive method that allows for detailed three-dimensional visualisation of the internal microstructure of materials. In the context of using phosphorus-rich residual streams in combustion for further ash recycling, physical properties of ash particles can play a crucial role in ensuring effective nutrient return and sustainable practices. In previous work, parameters such as surface area, porosity, and pore size distribution, were determined for ash particles. However, the image analysis involved binary segmentation followed by time-consuming manual corrections. The current work presents a method to implement deep learning segmentation and an approach for quantitative analysis of morphology, porosity, and internal microstructure. Deep learning segmentation was applied to microtomography data. The model, with U-Net architecture, was trained using manual input and algorithm prediction.•The trained and validated deep learning model could accurately segment material (ash) and air (pores and background) for these heterogeneous particles.•Quantitative analysis was performed for the segmented data on porosity, open pore volume, pore size distribution, sphericity, particle wall thickness and specific surface area.•Material features with similar intensities but different patterns, intensity variations in the background and artefacts could not be separated by manual segmentation – this challenge was resolved using the deep learning approach.

Small sized centroblasts as poor prognostic factor in follicular lymphoma - Based on artificial intelligence analysis

Histological assessment of centroblasts is an important evaluation in the diagnosis of follicular lymphoma, but there is substantial observer variation in assessment among hematopathologists.We aimed to perform quantitative morphological analysis of centroblasts in follicular lymphoma using new artificial intelligence technology in relation to the clinical prognosis. Hematoxylin and eosin slides of lesions were prepared from 36 cases of follicular lymphoma before initial chemotherapy. Cases were classified into three groups by clinical course after initial treatment. The ‘excellent prognosis’ group were without recurrence or progression of follicular lymphoma within 60 months, the ‘poor prognosis’ group were those that had relapse, exacerbation, or who died due to the follicular lymphoma within 60 months, and the ‘indeterminate prognosis’ group were those without recurrence or progression but before the passage of 60 months. We created whole slide images and image patches of hematoxylin and eosin sections for all cases. We designed an object detection model specialized for centroblasts by fine-tuning YOLOv5 and segmented all centroblasts in whole slide images. The morphological characteristics of centroblasts in relation to the clinical prognosis of follicular lymphoma were analyzed. Centroblasts in follicular lymphoma of the poor prognosis group were significantly smaller in nuclear size than those in follicular lymphoma of the excellent prognosis group in the following points: median of nuclear area (p = 0.013), long length (p = 0.042), short length (p = 0.007), nuclear area of top 10 % cells (p = 0.024) and short length of top 10 % cells (p = 0.020). Cases with a mean nuclear area of <55 μm2 had poorer event-free survival than those with a mean nuclear area of ≥55 μm2 (p < 0.0123). AI methodology is suggested to be able to surpass pathologist's observation in capturing morphological features. Small-sized centroblasts will likely become a new prognostic factor of follicular lymphoma.

Quantitative morphological analysis of Deinococcus radiodurans elucidates complex dose-dependent nucleoid condensation during recovery from ionizing radiation.

The extremophile Deinococcus radiodurans maintains a highly organized and condensed nucleoid as its default state, possibly contributing to its high tolerance to ionizing radiation (IR). Previous studies of the D. radiodurans nucleoid were limited by reliance on manual image annotation and qualitative metrics. Here, we introduce a high-throughput approach to quantify the geometric properties of cells and nucleoids using confocal microscopy, digital reconstructions of cells, and computational modeling. We utilize this novel approach to investigate the dynamic process of nucleoid condensation in response to IR stress. Our quantitative analysis reveals that at the population level, exposure to IR induced nucleoid compaction and decreased the size of D. radiodurans cells. Morphological analysis and clustering identified six distinct sub-populations across all tested experimental conditions. Results indicate that exposure to IR induced fractional redistributions of cells across sub-populations to exhibit morphologies associated with greater nucleoid condensation and decreased the abundance of sub-populations associated with cell division. Nucleoid-associated proteins (NAPs) may link nucleoid compaction and stress tolerance, but their roles in regulating compaction in D. radiodurans are unknown. Imaging of genomic mutants of known and suspected NAPs that contribute to nucleoid condensation found that deletion of nucleic acid-binding proteins, not previously described as NAPs, can remodel the nucleoid by driving condensation or decondensation in the absence of stress and that IR increased the abundance of these morphological states. Thus, our integrated analysis introduces a new methodology for studying environmental influences on bacterial nucleoids and provides an opportunity to further investigate potential regulators of nucleoid condensation.IMPORTANCEDeinococcus radiodurans, an extremophile known for its stress tolerance, constitutively maintains a highly condensed nucleoid. Qualitative studies have described nucleoid behavior under a variety of conditions. However, a lack of quantitative data regarding nucleoid organization and dynamics has limited our understanding of the regulatory mechanisms controlling nucleoid organization in D. radiodurans. Here, we introduce a quantitative approach that enables high-throughput quantitative measurements of subcellular spatial characteristics in bacterial cells. Applying this to wild-type or single-protein-deficient populations of D. radiodurans subjected to ionizing radiation, we identified significant stress-responsive changes in cell shape, nucleoid organization, and morphology. These findings highlight this methodology's adaptability and capacity for quantitatively analyzing the cellular response to stressors for screening cellular proteins involved in bacterial nucleoid organization.

The value of synthetic MRI in detecting the brain changes and hearing impairment of children with sensorineural hearing loss.

Sensorineural hearing loss (SNHL) can arise from a diverse range of congenital and acquired factors. Detecting it early is pivotal for nurturing speech, language, and cognitive development in children with SNHL. In our study, we utilized synthetic magnetic resonance imaging (SyMRI) to assess alterations in both gray and white matter within the brains of children affected by SNHL. The study encompassed both children diagnosed with SNHL and a control group of children with normal hearing {1.5-month-olds (n = 52) and 3-month-olds (n = 78)}. Participants were categorized based on their auditory brainstem response (ABR) threshold, delineated into normal, mild, moderate, and severe subgroups.Clinical parameters were included and assessed the correlation with SNHL. Quantitative analysis of brain morphology was conducted using SyMRI scans, yielding data on brain segmentation and relaxation time.Through both univariate and multivariate analyses, independent factors predictive of SNHL were identified. The efficacy of the prediction model was evaluated using receiver operating characteristic (ROC) curves, with visualization facilitated through the utilization of a nomogram. It's important to note that due to the constraints of our research, we worked with a relatively small sample size. Neonatal hyperbilirubinemia (NH) and children with inner ear malformation (IEM) were associated with the onset of SNHL both at 1.5 and 3-month groups. At 3-month group, the moderate and severe subgroups exhibited elevated quantitative T1 values in the inferior colliculus (IC), lateral lemniscus (LL), and middle cerebellar peduncle (MCP) compared to the normal group. Additionally, WMV, WMF, MYF, and MYV were significantly reduced relative to the normal group. Additionally, SNHL-children with IEM had high T1 values in IC, and LL and reduced WMV, WMF, MYV and MYF values as compared with SNHL-children without IEM at 3-month group. LL-T1 and WMF were independent risk factors associated with SNHL. Consequently, a prediction model was devised based on LL-T1 and WMF. ROC for training set, validation set and external set were 0.865, 0.806, and 0.736, respectively. The integration of T1 quantitative values and brain volume segmentation offers a valuable tool for tracking brain development in children affected by SNHL and assessing the progression of the condition's severity.

The trichothecene mycotoxin deoxynivalenol facilitates cell-to-cell invasion during wheat-tissue colonization by Fusarium graminearum.

Fusarium head blight disease on small-grain cereals is primarily caused by the ascomycete fungal pathogen Fusarium graminearum. Infection of floral spike tissues is characterized by the biosynthesis and secretion of potent trichothecene mycotoxins, of which deoxynivalenol (DON) is widely reported due to its negative impacts on grain quality and consumer safety. The TRI5 gene encodes an essential enzyme in the DON biosynthesis pathway and the single gene deletion mutant, ΔTri5, is widely reported to restrict disease progression to the inoculated spikelet. In this study, we present novel bioimaging evidence revealing that DON facilitates the traversal of the cell wall through plasmodesmata, a process essential for successful colonization of host tissue. Chemical complementation of ΔTri5 did not restore macro- or microscopic phenotypes, indicating that DON secretion is tightly regulated both spatially and temporally. A comparative qualitative and quantitative morphological cellular analysis revealed infections had no impact on plant cell wall thickness. Immunolabelling of callose at plasmodesmata during infection indicates that DON can increase deposits when applied exogenously but is reduced when F. graminearum hyphae are present. This study highlights the complexity of the interconnected roles of mycotoxin production, cell wall architecture and plasmodesmata in this highly specialized interaction.

Quantitative morphological analysis framework of infant cranial sutures and fontanelles based on CT images.

Characterizing the suture morphological variation is a crucial step to investigate the influence of sutures on infant head biomechanics. This study aimed to establish a comprehensive quantitative framework for accurately capturing the cranial suture and fontanelle morphologies in infants. A total of 69 CT scans of 2-4 month-old infant heads were segmented to identify semilandmarks at the borders of cranial sutures and fontanelles. Morphological characteristics, including length, width, sinuosity index (SI), and surface area, were measured. For this, an automatic method was developed to determine the junction points between sutures and fontanelles, and thin-plate-spline (TPS) was utilized for area calculation. Different dimensionality reduction methods were compared, including nonlinear and linear principal component analysis (PCA), as well as deep-learning-based variational autoencoder (VAE). Finally, the significance of various covariates was analyzed, and regression analysis was performed to establish a statistical model relating morphological parameters with global parameters. This study successfully developed a quantitative morphological framework and demonstrate its application in quantifying morphologies of infant sutures and fontanelles, which were shown to significantly relate to global parameters of cranial size, suture SI, and surface area for infants aged 2-4 months. The developed framework proved to be reliable and applicable in extracting infant suture morphology features from CT scans. The demonstrated application highlighted its potential to provide valuable insights into the morphologies of infant cranial sutures and fontanelles, aiding in the diagnosis of suture-related skull fractures. Infant suture, Infant fontanelle, Morphological variation, Morphology analysis framework, Statistical model.

Extensive palaeo-surfaces beneath the Evans–Rutford region of the West Antarctic Ice Sheet control modern and past ice flow

Abstract. The subglacial landscape of Antarctica records and influences the behaviour of its overlying ice sheet. However, in many places, the evolution of the landscape and its control on ice sheet behaviour have not been investigated in detail. Using recently released radio-echo sounding data, we investigate the subglacial landscape of the Evans–Rutford region of West Antarctica. Following quantitative analysis of the landscape morphology under ice-loaded and ice-unloaded conditions, we identify 10 flat surfaces distributed across the region. Across these 10 surfaces, we identify two distinct populations based on clustering of elevations, which potentially represent remnants of regionally coherent pre-glacial surfaces underlying the West Antarctic Ice Sheet (WAIS). The surfaces are bounded by deeply incised glacial troughs, some of which have potential tectonic controls. We assess two hypotheses for the evolution of the regional landscape: (1) passive-margin evolution associated with the break-up of the Gondwana supercontinent or (2) an extensive planation surface that may have been uplifted in association with either the West Antarctic Rift System or cessation of subduction at the base of the Antarctic Peninsula. We suggest that passive-margin evolution is the most likely of these two mechanisms, with the erosion of glacial troughs adjacent to, and incising, the flat surfaces likely having coincided with the growth of the WAIS. These flat surfaces also demonstrate similarities to other identified surfaces, indicating that a similar formational process may have been acting more widely around the Weddell Sea embayment. The subsequent fluctuations of ice flow, basal thermal regime, and erosion patterns of the WAIS are therefore controlled by the regional tectonic structures.

Morphometric evaluation of mitochondrial networks in regenerating myofibers using photothermal microscopy

Background: Mitochondria constitute their interconnected network through dynamic mitochondrial processes such as fission and fusion in response to changes in their intracellular environment, such as energy demand. Transmission electron microscopy is the standard method for evaluating mitochondrial morphology. We have developed a simple method to evaluate mitochondrial morphology using tissue sections for bright-field observation. In this study, we attempted to image light-absorbing molecules (cytochrome c) in mitochondria using photothermal (PT) microscopy. Purpose: This method can observe an entire myocyte, so this study attempted to evaluate the entire mitochondrial network of immature (regenerating) myocytes during the skeletal muscle regeneration process. Methods: The gastrocnemius muscle of anesthetized male Wistar rats (13 weeks old) was subjected to 300 controlled eccentric contractions (ECC) using electrical stimulation. 7 days after ECC loading, in situ perfusion fixation was performed, and transverse sections (1 μm) from red (deep regions) and white (superficial regions) were prepared after Epon embedding. The sections were observed by PT microscopy, and quantitative analysis of mitochondrial morphology was performed. Mitochondrial content was expressed as a percentage of the fiber total area in transverse sections. The PT imaging system uses lasers with wavelengths of 515 nm and 638 nm constituting the PT pump and the probe beams, respectively. Results: After 7 days of ECC, many myofibers with small fiber diameters and central nuclei, which are characteristic of the recovery process after muscle damage, were observed. The cross-sectional area of the regenerating muscle (superficial region 389.9 ± 15.2 μm2, deep region 366.2 ± 17.5 μm2) was significantly smaller than that of the normal muscle (superficial region 2211.1 ± 127.3 μm2, deep region 1396.1 ± 72.9 μm2). Mitochondrial content of the regenerating muscle (superficial region 26.1 ± 1.1%, deep region 22.7 ± 0.8%) was significantly lower than that of the normal muscle (superficial region 31.8 ± 1.9%, deep region 34.9 ± 1.8%). In normal muscle, a linear network of mitochondria along myofibrils was observed. On the other hand, no spatially distributed mitochondria were observed in the cytoplasm of the regenerating muscle. Furthermore, a mitochondrial network was formed around the periphery of the central nucleus. Conclusion: In muscle tissue imaging using PT microscopy, myofibers in the regenerative process have less mitochondrial content than mature muscle fibers and lack the mitochondrial network along myofibrils that characterizes mature muscle. On the other hand, the central nucleus of immature muscle was surrounded by highly developed mitochondria. The mitochondrial morphological characteristics of regenerating muscle fibers may offer a unique window through which to better understand mitochondrial design and function and their plasticity during maturation. This study was supported in part by a Grant-in-Aid for Japan Society for the Promotion of Science (JSPS) KAKENHI Grant (No. JP20H04074, 21K19703). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Quantitative Analysis of CGRP-IR Afferent Axons in the Mouse Stomach Using Zeiss Arivis Vision4D for Automated Tracing

Gastric nociceptive afferents detect and respond to noxious stimuli by transmitting signals from the stomach to the brain. Calcitonin gene-related peptide (CGRP) is a marker for nociceptive afferents and is crucial to gastric functions, including gastric acid secretion, gut motility, and inflammation. However, the topographical quantitative analysis of CGRP-immunoreactive (CGRP-IR) axons in the whole stomach is challenging due to the large amount of axon innervation in various targets within the tissue. Manual tracing of axons is time-consuming, labor-intensive, and extremely tedious, while currently existing automatic neuron tracing programs often need help with large file sizes or tracing inaccuracies. In this study, we used Zeiss Arivis Vision4D, a novel and sophisticated software with a user-guided machine learning capability, for automated tracing of CGRP-IR axons in flat-mounts of the whole mouse stomach muscular layers and thick serial sections of the antrum-pylorus region (C57BL/6J, male, 2-3 months old, n=7). We utilized a combination of techniques, including the Zeiss M2 Imager for automated scanning, Zen 3.3 (blue edition) for image processing, and Vision4D for stitching the entire stomach flat-mounts and sections, 3D fiber tracing, and density analysis. Our results demonstrated that: 1) Vision4D detected and traced almost all CGRP-IR axons of different gastric targets (blood vessels, muscles, myenteric neurons) in different regions (fundus, corpus, antrum) of the stomach. 2) The x-y-z tracing allowed the visualization of axons in space, portraying its continuous trajectory across layers in the muscular flat mounts (longitudinal muscle, myenteric plexus, circular muscle) and cross sections (muscles, submucosa, mucosa). 3) Vision4D also enabled quantitative morphological and laminar analysis based on target and layer separations. The axon density heatmap generated from the tracing data showed high CGRP-IR axon innervation in the blood vessels. After the removal of blood vessels, dense innervation was seen in the upper fundus and antrum-pylorus followed by corpus, indicating the abundance of CGRP in these regions. 4) Vision4D is a powerful tool in handling large data sizes (hundreds of gigabytes of optical sections in more than 300 image frames per stomach) and outstanding in its capability to track nerve fibers in a highly accurate and time effcient manner. This has paved the way for data batch processing to be accessible, as well as shown that multi-marker tracing of the same tissue sample is attainable. 5) The tracing data will be integrated into a stomach scaffold for a 3D map of CGRP-IR afferent axons. This study represents, for the first time, the automatically traced CGRP-IR axons using whole stomach preparations and highlights the potential of Vision4D in creating axon tracing models in other organs and species. This study is supported by NIH 1 U01 NS113867-01, NIH R15 1R15HL137143-01A1 and Carl ZEISS Company. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.