Modern microscopy systems allow researchers to generate large volumes of image data with relative ease. However, the challenge of analyzing this data effectively is often hindered by a lack of computational skills. This bottleneck negatively impacts both research reproducibility and efficiency, as researchers frequently rely on manual or semi-automated analysis methods. Interactive image analysis workshops offer a valuable solution, equipping researchers with the skills and tools needed to automate image processing tasks. In this paper, we share our experiences and best practices from conducting such workshops, which emphasize the use of open-source software like ImageJ, FIJI, and Python-based tools such as JupyterLab and napari. We discuss key considerations for workshop design, logistics, and outcomes, while highlighting common pitfalls to avoid. Using two recent workshops as case studies, we also present strategies for optimizing participant engagement and learning. Our insights offer practical guidance for planning and conducting image analysis workshops and serve as a starting point for researchers looking to establish similar training initiatives and enrich their local imaging communities.

Quinone outside inhibitor (QoI) fungicide resistance in A. alternata populations was reported in Brazil for the first time in 2019, in São Paulo orchards, and the mutation G143A in cytochrome b ( cytb) was found in resistant isolates. Our study investigated the infectious process, production of Reactive Oxygen Species (ROS) and fungal cell death in resistant (QoI-R) and sensitive (QoI-S) Alternaria alternata pathotype tangerine (Aapt) isolates. Morphological characterization of Aapt isolates was performed using Confocal Laser Scanning Microscopy (CLSM). Alternaria brown spot (ABS) symptoms were produced by Aapt isolates on tangelo cv. BRS Piemonte. Germination of QoI-R conidia and production of germ tubes on tangelo leaflets treated with 100µg mL -1 of pyraclostrobin 18 hours after inoculation (hai) was observed using Scanning Electron Microscopy (SEM). At the same time, QoI-S conidial germination was inhibited on tangelo leaflets treated with pyraclostrobin. ROS production and cell death in Aapt isolates at high fungicide concentration were observed using CLSM. QoI-S conidia exhibited high ROS production, indicating high oxidative stress. When dyed with Propidium Iodate (PI), QoI-S conidia emitted red fluorescence, showing cell death and confirming their sensitive phenotype. In contrast, QoI-R conidia neither produced ROS nor exhibited red fluorescence, indicating no cell death and confirming their resistant phenotype. Therefore, our findings evidence that microscopic techniques may help characterize events during fungi-plant interactions, ROS production, cell death, and Aapt phenotypes resistant and sensitive to QoIs using fluorometric protocols. Research Highlights Our findings demonstrate the infection process, reactive oxygen species production, and cell death of Aapt isolates resistant and sensitive to quinone outside inhibitors (QoIs). Confocal laser scanning microscopy may help to characterize phenotypes resistant and sensitive to QoIs using fluorometric protocols.

Waveguide evanescent field fluorescence microscopy (WEFF) is an evanescent based microscopy utilizes a confined thin film of light, around 100 nm, to image the plasma membrane of cells attached to a waveguide. Low photobleaching and low background beside its high axial resolution allows time-lapse imaging to investigate changes in cell morphology in the presence or absence of chemical agents. Both large field of view (FOV) and uniform illumination are very important while imaging cell-substrate contacts with an evanescent field. In the current work, we demonstrate that the WEFF microscope is capable of large FOVs with a uniform illumination source and imaging over a very long time period with a simple and an inexpensive experimental setup. The interaction of the trypsin with plasma membranes of live osteoblast cells is investigated. To analyze cell images (250 images), instead of relying on manual tracking, which is time-consuming and can introduce numerous errors, we performed image processing using TrackMate to investigate the dynamic response of cells upon exposure to trypsin. This helps to save time and increase the accuracy of the analysis. The powerful tracking and analysis capabilities of the TrackMate plugin in ImageJ is used to automatically detect the cells boarder and traces each cluster of cells. The reduction in cell area is accompanied by a notable increase in mean intensity, reflecting changes in the intracellular environment. However, the background did not change during the experiment, which proves that the fluorescence material remains attached to the cell membrane and does not leak into the cell medium.

The Role of the Ocular Lids of the Black-Winged Kite, Elanus caeruleus, in the Immune Protection of the EyeNahed Ahmed Shawki1, Abdelraheim H Attaai2, Amany Mohamed Abdel- Mageed1, Fatma Abdel- Regal Mahmoud11 Assiut University, Faculty of Science. Zoology Department, Assiut, Egypt.2 Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Assiut University, Assiut, Assiut 71526, Egypt.2 Department of Anatomy and Histology, School of Veterinary Medicine, Badr University in Assiut, Assiut 11829, Egypt; [email protected]

Skeletal porous lesions such as cribra orbitalia (CO) have long been of interest to bioanthropologists worldwide, mainly due to their high prevalence in osteological material. Previous studies considered CO as an external morphological manifestation, and therefore, research has mainly focused on visible (macroscopic) CO patterns. However, the understanding of CO-induced micro-scale bone changes is still scarce. Therefore, here we performed high-resolution micro-computed tomography imaging to investigate three-dimensional CO-induced micro-architectural patterns in non-adults, with a particular focus on the correlation between macroscopic and micro-architectural orbital features. Cortical and trabecular micro-architectural changes in the orbital roof were analyzed in non-adults up to 15 years of age, using orbital roof samples with and without macroscopic traces of CO (n=28). A widely accepted five-grade macroscopic CO scoring system was applied to analyze CO severity. Areas affected with CO (area 1) and areas without macroscopic CO traces (area 2) were analyzed separately. The conducted high-resolution analysis showed that cortical and trabecular micro-architecture varied with CO presence, lesion severity (CO grade), and the analyzed area. Inter-grade comparisons suggested that most of the analyzed micro-architectural parameters were not significantly different between adjacent CO grades. Based on the micro-architectural evaluation of areas 1 and 2, the porous lesions were much more extensive than revealed by gross examination. Our analysis also revealed that micro-architectural differences were particularly pronounced in younger non-adults. Taken together, the macroscopic examination of CO appears to reflect only the tip of the iceberg, as the micro-architectural changes are much larger than macroscopically identified.

We present the development of a custom-built structured illumination microscope (SIM) featuring a specially designed transmission diffraction grating. Employing the analogue microfilming method, we fabricated transmission diffraction gratings suited to the specific requirements of our system. This robust and cost-effective method allows for the fabrication of diffraction gratings with customised constants, ensuring excellent transmission in both the visible and near-infrared spectra. Additionally, to assess the performance of our system, we measured the resolution in both epifluorescent and superresolution imaging modalities by applying two independent methods: the conventional resolution measurement using fluorescent beads and the knife-edge technique applied on the MoS 2 monolayer flakes. Both methods confirmed enhancement in the resolution of SIM over the epifluorescent imaging modality. Furthermore, we have successfully demonstrated the capabilities of our microscope by imaging fluorescently labelled astrocytes, specifically targeting the vimentin filament protein within these cells. The superresolution images reveal fine structures of the vimentin cytoskeleton that remain unresolved in the epifluorescent image.

This study was carried out to investigate the macro-anatomical, morphometric, histological and electron microscopic characteristics of the uropygial gland in adult male and female turkeys (Meleagris Gallapova). The glandula uropygialis of thirteen adult turkeys (6 males, 7 females) were used for this study. It was determined that this glandular structure located dorsal to the last caudal vertebrae was heart-shaped and consisted of two lobes and a papilla system. As a result of statistical evaluation, significant differences were found between the sexes in the parameters of lobe length, papilla width and papilla height (P<0.05). Also, PUI value had a statistically significant difference (P<0.05). Tissue samples taken to determine the histological structure of the gland were stained with Haematoxylin & Eosin, Crossman’s and Periodic Acid Schiff. Histological examination revealed that the gland had a two-lobed structure surrounded by a capsule composed of connective tissue. It was determined that the gland had a tubuloalveolar-holocrine structure and the epithelial layer consisted of cellular layers as germinative layer, intermediate layer, secretory layer and degenerative layer from the periphery to the centre. In the scanning electron microscope examination, the lobe structure of the gland consisted of different shaped follicles protruding from the surface. It was observed that these glandular follicle structures were gathered together in different ways or mostly disorganised. As a result of the study, it was determined that the morphological and histological structure of the uropygial gland in turkeys was similar to that of other bird species, but showed some species-specific and habitat-dependent differences in general. Research Highlights: The uropygial gland consisted of a system of two lobes and one papilla in each of male and female turkeys. The glandular lobes consisted of follicles surrounded by connective tissue, while the follicles were connected to each other by interfollicular septae. The results obtained in morphometric measurements revealed statistical differences between male and female birds. Histological examination showed PAS (+) reaction in the basal membranes of tubule epithelial cells and secretion. Lymphoid cell communities were found in both connective tissue and intertubular regions.

Detecting RNA molecules within their natural environment inside intact arthropods has long been challenging, particularly in small organisms covered by a tanned and pigmented cuticle. Here, we have developed a methodology that enables high-resolution analysis of the spatial distribution of transcripts of interest without having to dissect tiny organs or tissues, thereby preserving their integrity. We have combined an in situ amplification approach based on Hybridization Chain Reaction, which enhances the signal-to-noise ratio, and a clearing approach that allows the visualization of inner organs beneath the cuticle. We have implemented this methodology for the first time in Hemiptera, mapping two salivary aphid ( Acyrthosiphon pisum) transcripts, the effector c002 and the salivary sheath protein SHP. With a multiplex approach, we could simultaneously detect different mRNAs in whole-mount pea aphid head-thorax samples and show that they were distributed in distinct secretory cells of salivary glands.

In this article we present a new method called PSF-Radon transform algorithm. This algorithm consists on recovering the instrument point spread function (PSF) from the Radon tranform (in the line direction axis) of the Line Spread function (that is, the image of a line). We present the method and tested with synthetic images, and real images from macro lens camera and microscopy. A stand-alone program along with a tutorial is available, for any interested user, in [? ] .

Here, we describe for the first time the sperm morphology of Tingidae (Heteroptera). They are small insects presenting lacy patterns on their pronotum and hemielytra and are exclusively phytophagous, with many economically important species. We studied five species of the tribe Tingini (Tinginae): Teleonemia scrupulosa, Vatiga illudens, Gargaphia lunulata, Leptopharsa sp., and Corythucha arcuata. Their spermiogenesis process is similar to other Heteroptera, with some differences in the formation of the centriole adjunct. This structure extends in the anteroposterior spermatid axis, flanking the nucleus, possibly contributing to nucleus remodelling and sperm elongation. The mature sperm of Tingidae is also similar to that of other Heteroptera, with features that corroborate the group’s monophyly. Our data support previous results for their sister family, Miridae, which exhibits some characteristics exclusive to this taxon, not present in Tingidae or other Heteroptera. They also support the sister relationship of the genera Gargaphia and Leptopharsa and suggest closer relationship between Vatiga and Corythucha. Overall, this study sheds light on the sperm ultrastructure of Tingidae and provides information for understanding the evolution and diversity of Heteroptera.

Recent advances in computing power triggered the use of Artificial Intelligence in image analysis in life sciences. To train these algorithms, a large enough set of certified labelled data is required. The trained neural network is then capable of producing accurate instance segmentation results, that will then need to be re-assembled into the original dataset: the entire process requires substantial expertise and time to achieve quantifiable results. To speed-up the process, from cell organelle detection to quantification across modalities, we propose a deep learning based approach for Fast AutoMatic Outline Segmentation (FAMOUS), that involves organelle detection combined with image morphology, and 3D meshing to automatically segment, visualize and quantify cell organelles within volume electron microscopy datasets. From start to finish, FAMOUS provides full segmentation results within a week on previously unseen datasets. FAMOUS was showcased on a dataset acquired using a focused ion beam scanning electron microscope (FIBSEM), and on yeast cells acquired by transmission electron microscopy.

To detect hydrogen in materials at the atomic scale, atom probe tomography is now regularly used. In order to avoid cumbersome cryo-preparation to suppress diffusion, often hydrogen is charged only into the finished specimen. Here, the use of hydrogen gas over electrochemical hydrogen has the advantage that the specimen is not contaminated with an electrolyte. So far, this ‘charging’ has been done in large, expensive systems. Here, we introduce small devices that enable the exposure of atom probe specimens to hydrogen and potentially other gases, using only very small gas volumes. This enables the operation in regular laboratory environments without additional safety measures. These devices can be used to expose the specimen to hydrogen up to 10 bar / 90 °C. Higher temperatures may be attained with small changes. Validation of the success of charging with these setups is demonstrated through experiments employing deuterium charging of palladium atom probe tips.

Beetle hindwings have the unique advantages of lightweight and high strength, which play a key role in flight. In this study, the beetle hindwings were cut along the chordal direction, then the first groove microstructure of different vein cross sections was investigated using the 3D microscope system and LSCM. It was found that the position of the first groove relative to the entire chordal cross-section of the wing gradually moves backwards, which has an effect on the flying aerodynamic behaviors of the beetle. Next, three corrugated airfoils (CA models) learned from the microscopy imaging of ladybird beetle hindwing were designed. Then aerodynamic behaviors were calculated by ANSYS Fluent software, it was confirmed that the position of the first groove microstructure affects the aerodynamic performance of the airfoil. For further study the influence of corrugated structural and motion parameters on the aerodynamic, 2D ‘simplified’ airfoil models with triangular corrugations (TWA models) were developed and studied.

This is a very first attempt to study various parameters of a medicinal plant, Delphinium suave Huth. The plant is erect, geophytic, herbaceous, with tuberous root, trifid in a palmatipartite, strigose cuneate leaf and white spurred zygomorphic flower. The root was isodiametric phellem with single non-glandular trichomes. The stem revealed single-layered cuticle, multiseriate epidermis, cortex, pith ray and uniserate bowed non-glandular trichomes. The leaf was amphistomatic, showed tapering trichomes, prismatic crystals and ranunculaceous stomata with circumference 144.66-182.67 µm. Pollen grains in Light Microscopy (LM), were prolate, spheroidal trizonocolpate, isopolar, radiosymmetric, scabrate, elliptic and monads. Scanning Electron Microscope (SEM) pollen surface was scabrate, monad, size varied from 18.06 µm-16.67µm, colpus to inaperturate, tricolpate, ornamented, echinus, isopolar, isodiametric and circular. SEM roots showed sclerenchymatic tissues, stellate, glandular, non-glandular trichomes and crystals. The stem showed scalariform, pitted vessels, warty protuberances, unicellular, silicified, non-glandular trichomes. Leaves powder revealed, simple, unicellular, tapered headed, uniseriate, sessile, capitate, unbranched glandular, non-glandular, trichomes with crystals. Capitate, stellate, circular, unicellular, branchy trichomes were observed for the first time through SEM. Powder drug study of root, stem leaves through LM also revealed different tissues. Preliminary phytochemical revealed alkaloids, anthocyanins, anthraquinones, coumarins, flavones, mucilages, saponins, steroids, terpenoids, volatile oils and proteins. GC/MS showed 36 compounds in roots 33 in stem while 40 in leaves. Fluorescence analysis of roots, stem and leaves showed variation in colour when treated with chemicals. This study will assist pharmacognostic exploration, authentication from adulterants/allied species for consistent quality, resulting in safe use, preservation and efficacy.

ABSTRACT The double-spined spruce bark beetle, Ips duplicatus has become an infamous secondary pest of Norway spruce, causing extensive ecological and economic destruction in many Central European countries. Antennae are the primary olfactory organs that play fundamental role in insect-host chemical communication; therefore, understanding morphology is crucial before conducting electrophysiological investigations. Here, we present our analysis of sensilla types on the antennal surface of I. duplicatus for the first time, using high-resolution-scanning electron microscopy. We studied the external morphological characteristics of antennae and the types, numbers, and distribution of the antennal sensilla in males and females. Our results revealed the presence of five different types of morphologically distinct sensilla: sensilla chaetica, sensilla basiconica, sensilla trichodea, sensilla coeloconica, and Bӧhm’s bristles. We observed two subtypes in sensilla chaetica (SChI and SChII), four subtypes in sensilla basiconica (SBI, SBII, SBIII and SBIV), three subtypes in sensilla trichodea (STrII, STrIII and STrIV) and two subtypes in sensilla coeloconica (SCoI and SCoII), respectively in I. duplicatus males and females. Minor differences in length and numbers between the sexes for some sensilla types were found. Distribution maps for different sensillar types were constructed and specific areas for the respective sensilla were found. Possible functions of observed sensilla types are discussed. The present study provides a basis for future electrophysiological studies aimed at understanding how I. duplicatus detects ecologically important olfactory cues.

Aim: This study aimed to use the micro-computed tomography to evaluate the interfacial adaptation and the presence of gaps of NeoMTA Plus, BioRoot RCS, and MTA in the root-end cavities. Methodology: Thirty standardized bovine roots measuring 15 mm in length were selected. Chemical-mechanical preparation was performed up to instrument #80 and obturation with the cold lateral compaction technique with cement based on zinc oxide and eugenol. The roots were kept at 37º C for seven days. Afterward, apicectomy of the apical 3mm and a root-end filling cavity was performed at 3mm depth. Micro-computed tomography (micro-CT) was performed to measure the volume of the retroactivity. The roots were divided by stratified randomization into three groups according to the retro-end filling material: NeoMTA Plus, BioRoot RCS, and MTA. A new micro-CT was performed to assess the presence of voids in the root-end filling material and between it and the canal wall. One-way ANOVA and Tukey tests were performed using the BioEstat 4.0 program. Results: There was no difference in the initial volume values of the root-end cavities (P > 0.05). After the insertion of root-end filling materials, the most significant volumes of voids were observed in the NeoMTA Plus group (P < 0.05), with no difference for the BioRoot RCS and MTA Angelus groups (P > 0.05). Conclusion: Micro-computed tomography showed that MTA and BioRoot RCS have better interfacial adaptation and presented fewer number of gaps than NeoMTA Plus when used as root-end filling materials.

We investigated the gross and ultrastructure morphology of the mandibles, labial palpus and mandibular palpus of adult male and female Glenea cantor beetles using scanning electron microscopy. There were significant differences between males and females in the gross and ultrastructure morphology of these parts. Observation of the gross morphology of the mandible, labial palpus and mandibular palpus revealed that their lengths were significantly greater in females, which is mainly related to the oviposition mechanism. In terms of the ultrastructural morphology of the mandibles, labial palpus and mandibular palpus, seven types of sensilla were found on these parts: sensilla twig basiconica (STB I, II, III, and IV), sensilla chaetica (SCh IV and V), Böhm’s bristles (Bb), sensilla placodea (SP), sensilla trichodea (ST II), sensilla plate (SP), sensilla coeloconica (SC) and sensilla campaniformia (SCa). Females have significantly more ST I and SCh III on their mandibles than males. The mechanical sensors SCh and Bb, olfactory sensor ST II, taste sensor STB IV, and carbon dioxide sensor and temperature and humidity sensors SC, SP and SCa were significantly more developed on female than male labial and maxillary palps, which also further explained the important role of the oral appendages in oviposition behavior. These results will help us further understand the host selection and oviposition behavior of adult G. cantor and promote the information-based design of pest control methods.

Sperm morphology is considered a species-specific character and has been used as a tool in the classification of numerous mammalian taxa. Neotropical bats have been poorly studied, and important aspects on sperm morphology have not been elucidated. The aim of the present study was to describe and compare the sperm morphology and morphometry of Molossus molossus and Molossops temminckii. 14 adults specimens were analyzed from the Colección Mamíferos Lillo, Universidad Nacional de Tucumán: five Molossus molossus and nine Molossops temminckii. The epididymis were extracted and macerated in Farmer’s solution, followed by a coloration with different stains. To carry out the description and morphometric analysis, microphotographs were taken under an optical, epifluorescence and scanning electron microscope. A total of 50 sperm from each individual were measured for morphometric analysis. The length and width of the head, midpiece and tail were taken as variables. Sperm from M. molossus and M. temminckii were practically identical, both morphologically and morphometrically. In both species, a distal bulge was observed at the end of the intermediate piece in a percentage greater than 85%. The main characteristics shared between the species were: presence of acrosomal blebs in the upper half of the head of the spermatozoa; cephalic equatorial segment with filiform ornamentations; intermembrane space of head apex wedge-shaped; helical middle piece and annulus at the end of middle piece. In the present study, scanning electron microscopy allowed us to visualize structures, such as acrosomal vesicles, that were not detected with other types of microscopy.