Digital Microscopy Research Articles

Response Time Dynamics of a Membrane-Based Microfluidic Gas Sensor

Practical gas–liquid interfacing is paramount in microfluidic technology, particularly in developing microfluidic gas sensors. We have created an easily replicable membrane-based closed microfluidic platform (MB-MP) to achieve in situ gas–liquid contact for low-resource settings. We have fabricated the MB-MP using readily available materials like double-sided tape or parafilm without conventional soft lithographic techniques. The response characteristics of the MB-MP are studied using CO2 as the model gas and bromothymol blue dye as the sensing material. The dye’s color change, indicative of pH shifts due to CO2 absorption, is captured with a digital microscope and analyzed via the ImageJ software package v1.54g. The response shows saturation and regeneration parts when cycled between CO2 and N2, respectively. Experiments are conducted to investigate the response characteristics and saturation rate under different conditions, including changes in volumetric flow rate, gas stream velocity, and dye solution volume. We observe experimentally that an increase in volumetric flow rate decreases the delay and increases the saturation rate of the response, surpassing the impact of the gas stream’s increased velocity. Furthermore, increasing the dye volume results in an exponential decrease in the saturation rate and an increase in the delay. These insights are essential for optimizing the platform’s response for point-of-use applications.

Retention and wear assessment of two different bar materials used in implant-retained mandibular overdenture

Abstract Aim To assess retention and wear of two different bar materials (cobalt chromium and poly ether-ether ketone) used in implant-retained mandibular overdenture subjected to chewing simulation Robota during 1 year of functional simulation. Patients and methods This study is a comparative laboratory study which was carried out on 14 models of implant-retained mandibular overdenture, for each model two implants were inserted. The 14 models were divided equally into two equal groups: group A (overdenture constructed over two implant analogs with bar attachment and clip made from cobalt chromium material), and group B (overdenture constructed over two implant analogs with bar attachment and clip made from poly ether-ether ketone material). Drilling of the models was done using milling machine and each model received two implant analogs inserted 11 mm from midline at canine region. The overdentures were constructed in the usual manners and the teeth were set in lingualized occlusion. The mandibular overdentures were subjected to 150 000 chewing cycles using Robota chewing simulator and 1080 insertion-removal cycles. Retention of the samples was measured by the universal testing machine and wear observed under USB digital microscope both of which were done at the beginning of the study and after 3, 6, 9, and 12 months. Results The retention force for group A decreased significantly from baseline 43.43 ± 5.25 to 35.56 ± 8.30 after 3 months of cyclic loading, with similar reductions after 6, 9, and 12 months. Group B also showed reductions from baseline 26.75 ± 2.32 to 21.29 ± 6.74 after 3 months, and similar patterns over time. Group A had no statistically significant wear changes after cyclic loading, with a baseline mean of 0.2904 ± 0.0008, changing to 0.2913 ± 0.0008 after 3 months, while group B showed similar nonsignificant changes. There were no statistically significant differences in wear between both groups at any time point. Conclusion Retentive forces decrease with increasing cyclic loading. However, both materials displayed acceptable retentive values. Wear values were nonsignificant between two studied groups.

First-ever study uncovers microplastic contamination in Nepalese table salt

Despite numerous studies on microplastic contamination in table salt worldwide, research focusing on Nepalese table salts is remarkably lacking. This study aims to address this gap by investigating microplastic contamination in salt samples collected from all seven provincial zones of Nepal. Our objective is to comprehensively assess the presence and characteristics of microplastics in salt sold within local markets across the country. Five salt packaging companies utilized by Nepalese consumers were identified. The collected salt samples were digested with Fenton's reagent to extract microplastics, which were then observed under a digital microscope. Using an OMAX stereomicroscope at 30× magnification and an OMAX A3503S digital camera, each microplastic was carefully identified and quantified. FTIR analysis was conducted to identify the polymer types. All tested salt samples (100 %) from both Nepalese and Indian packaging companies contained microplastics. Microplastic abundance exhibits variability among the samples, ranging from 80 to 1040 microplastics per kilogram of salt. The average value stands at 381 ± 219 microplastics per kilogram of salt sample. The distribution of microplastic concentrations within the salt samples reveals that the majority fall within the 301–400 microplastics per kilogram salt range, constituting 33 % of the total samples. Color analysis showed diverse contamination sources, while microplastic shapes included fibers (56 %), films (17 %), fragments (16 %) and pellets (11 %). Polymer type analysis confirmed the presence of polyethylene and polypropylene in tested microplastics. The study estimated that Nepalese individuals ingest an average of 1853 microplastics annually, indicating significant exposure from salt consumption. Surprisingly, factors such as storage conditions, date of salt packaging and thickness of packaging material did not significantly affect microplastic presence in the samples, suspecting manufacturing processes as the primary contributors to contamination. Therefore, the raw materials and purification practices for salt need improvement, as existing methods are insufficient to eliminate microplastics. These findings emphasize the need for further research and mitigation efforts to address microplastic contamination in Nepalese salt and its potential health impacts.

Comparative evaluation of shear bond strength of orthodontic brackets bonded with two self-etching adhesive systems: an in vitro study

Abstract Objective To evaluate and compare the shear bond strength (SBS of the new Optibond eXTRa Universal and Adper Prompt L-Pop self-etch adhesive systems using an in vitro experimental design. Patients and methods The sample included 40 freshly extracted human maxillary first premolars collected from orthodontic patients, adhering to ethical guidelines. The teeth were randomly assigned to two groups, one treated with Optibond eXTRa Universal self-etch adhesive and the other with Adper Prompt L-Pop. Stainless-steel brackets were then bonded to the teeth using BracePaste orthodontic adhesive composite resin. After storage and thermocycling, SBS was measured using a universal testing machine, and adhesive remnant index (ARI) scores were assessed under a digital microscope. Statistical analyses were performed, including the Shapiro–Wilk method, Student’s t-test for SBS, and χ 2 test for ARI scores. Results The mean SBS showed no significant difference between both groups (P = 0.076). However, the distribution of ARI scores exhibited a highly significant difference between the two groups (P < 0.001). Conclusion While both adhesives exhibited similar bond strengths, variations in adhesive residue patterns highlight the importance of considering adhesive performance in orthodontic applications such as ease of cleanup and potential impact on enamel health.

Titanium-Titanium Junctions in the Knee Corrode, Generating Damage Similar to the Hip

BackgroundPrevious studies identified corrosion between the modular tibial components of total knee arthroplasty devices. However, gaps persist. Compared to the hip, damage modes that occur within taper junctions in the knee remain poorly understood. In this study, we investigated corrosion on total knee arthroplasty components with titanium-titanium junctions. We asked the following question: under typical in vivo cyclic loading conditions, will the same alloy damage modes from total knee arthroplasty devices resemble those documented in the hip? MethodsA total of 50 paired titanium alloy tibial baseplates and stems were collected and semiquantitatively analyzed using Goldberg corrosion scoring. To characterize damage, a subsection of moderately and severely corroded components was sectioned and imaged using scanning electron and digital optical microscopy. ResultsOf the 100 device components, 95% showed visual evidence of corrosion. The initial contact area between the stem and bore generally occurred 3 mm from the stem taper base. Scanning electron microscopy revealed 4 damage modes, including oxide film formation, crevice corrosion, selective dissolution, and pitting. ConclusionsEach of the damage modes identified in modular titanium-titanium tibial junctions was previously reported by total hip arthroplasty retrieval studies. Cumulatively, our results suggest that mechanically assisted crevice corrosion promoted this damage in vivo.

A pilot study of dimeticone oils versus sodium carbonate treatment for tungiasis: A randomized cohort trial in Homa Bay County, Kenya.

Tungiasis is a cutaneous parasitosis caused by the female flea Tunga penetrans. Two-component dimeticone (NYDA) is the only treatment for tungiasis recommended by the World Health Organization; however, this topical drug is not available in Kenya. In Western Kenya, sodium carbonate is commonly used in the treatment of tungiasis. This study evaluated the 7-day cure rates for tungiasis by comparing sodium carbonate and NYDA treatments in Homa Bay County, Kenya. This was a randomized, observer-blinded, parallel-treatment cohort trial. Twenty-three eligible children with 126 flea infections were matched and randomized. All participants received both treatments, with one treatment on each foot. We recorded all health conditions/information, including inflammation scores and adverse events. Observations were performed on days 3, 5, and 7 using a digital microscope to confirm dead or live fleas based on the viability signs. Twenty-three children aged 3-13 years were analyzed. The proportion of dead fleas on day 7 was higher after NYDA treatment than after 5% sodium carbonate treatment (87% versus 64%, respectively, P = 0.01) NYDA. Median survival was 5 days for both treatments; NYDA had significantly higher trend of flea non-viability rate than 5% sodium carbonate (P<0.01). There were no significant differences in the inflammation score or pain/itchiness between the two treatments. On the last day, 14 children indicated their preference for NYDA in future treatment of tungiasis, whereas nine children preferred the 5% sodium carbonate solution. NYDA was significantly more effective than 5% sodium carbonate for tungiasis treatment. Both treatments were safe but the children preferred NYDA more. Future studies with more participants and an extended observation period are warranted to confirm our findings. The findings suggest that NYDA should be made more available in tungiasis endemic area. UMIN-CTR; UMIN 000044320.

A Comparative Evaluation of Marginal and Internal Fitness of Zirconia Monolithic Crown Fabricated with Different CAD\CAM Systems: (In Vitro Study)

The precision of computer-aided design/computer-aided manufacturing (CAD/CAM) systems is reliant upon their technical attributes and reliability in producing the designated restorations. The evaluation of this precision is primarily conducted through the examination of the marginal and internal fitness of said restorations. Consequently, the objective of this research endeavor was to analyze and compare the marginal and internal fitness of zirconia crowns fabricated using various CAD/CAM systems. The samples were categorized into four groups, each consisting of 10 samples, based on the specific CAD/CAM equipment employed for crown fabrication as follows: Group I fabricated using Sirona CAD\CAM milling machine, Group II fabricated using Dentium CAD\CAM machine, Group III fabricated using XTCERA CAD\CAM machine and Group IV fabricated using IMES-ICORE CAD\CAM machine. Tooth preparation done following the guidelines recommended by KATANA TM Zirconia (Kurary Noritake Dental Inc., Japan). To ensure standardization, one CAD/CAM program designed parameters was selected to be used for all CAD/CAM machines. Cement thickness was directly measured using sectioning technique to assess the crown's internal and marginal fitness. After performing sectioning, measurements were conducted using a digital microscope with a magnification of 230x. Eleven predetermined measuring locations were selected for each sample, representing four distinct areas: two marginal points, two chamfer points, four axial points, and three occlusal points. Results showed that none of four CAD\CAM machines tested was able to accurately reproduce the designed cement space parameter except Group I (Sirona) showed the closest results. As a conclusion, no CAD\CAM system is 100% accurate however, crowns fabricated using Sirona CAD\CAM milling machine showed the closest recordings to selected parameters among other CAD\CAM milling machines tested.

Full-Field Strain and Failure Analysis of Titanium Alloy Diamond Lattice

The advancement in additive manufacturing has significantly expanded the use of lattice structures in many engineering fields. Titanium diamond lattice structures, produced by a direct metal laser sintering process, were experimentally investigated. Two cell sizes were selected at five different relative densities. Morphological analysis was conducted by digital microscopy. The compressive tests and digital image correlation technique allowed the evaluation of elastic moduli to be used in the Gibson–Ashby model. Failure mechanisms of the structures have been analysed by digital image correlation, which represents a promising technique for strain evaluation of such structures. A non-linear finite element model of the lattice structures was developed and validated using the experimental data. The analysis of the results highlights the good mechanical properties of the Ti6Al4V alloy lattice structures.

Development of a digital holographic microscope model for the investigate of structures in the optical range

One of the modern and relevant methods for investigating the structures of objects is based on the holographic method of recording signals (holographic microscopy). The main advantage of this method is the ability to obtain complete information about the object. In other words, this method makes it possible to record not only the amplitude, but also the phase of the wave. This is achieved thanks to a recording scheme in which the phase of the wave is some modulation of the intensity. This advantage makes holographic microscopy an effective tool for the investigate of particles/microparticles in gases, liquids and solid materials in the form of thin films or in sufficiently transparent materials for optical waves (one of the main limitations of the holographic recording scheme is to investigate only objects with high transmissivity, i.e. the reference wave is must be about 70% or more of the total wave). Within the framework of this work, we consider the scheme of in-line holography (Gabor holography). The undoubted advantage of the in-line holographic investigation method is that it is limited only by the wavelength range. In other words, by changing the wavelength of the source, a wide range of objects can be examined. For example, in-line holography is used in low energy electron microscopes, which allows the atomic structure of an object to be studied. In the case when the source is a laser (optical range), a holographic microscope provides a wide range of possibilities for investigation the micro-objects, from various bacteria to various fine–structured particles. We developed a model of a digital holographic microscope for the study of structures in the optical range, based on the Gabor in-line holography method. This model of the microscope is developed on the Raspberry Pi Zero 2W platform.

Karakterisasi Tebal Lapisan Dan Ketahanan Korosi Mild Steel SS400 JIS G3101 Dengan Metode Elektroplating Ni-Cr

Mild steel is steel with a concentration of carbon element below 0.3%. In its use as an oil storage tank material on fuel transport trucks, there is a problem, namely its resistance to corrosion in fuel. Sulfur compounds in fuel have a role in the formation of corrosion. Currently, the Ni-Cr electroplating coating method is very widely used as an effort to control the corrosion rate of carbon steel. The use of Ni-Cr as an anode is relied upon as a substance resistant to corrosion. Therefore, the purpose of this study was to identify how layer thickness affects corrosion levels in SS400 mild steel based on current variations and duration of the electroplating process. In this study using mild steel SS400 JIS G3101 with electroplating method (electroplating) varying the electric current 1A, 2A and 3A, as well as electroplating time of 10 minutes, 20 minutes and 30 minutes. The test parameters of the electroplating results are the actual layer thickness test, theoretical thickness test and corrosion rate test. Data processing was analyzed using two-way variance analysis. The thickness of the layer increases with increasing electric current and the length of time of Ni-Cr electroplating. The highest thickness value of 102.34 μm using a digital microscope while for theoretical testing of 98.33 μm, both were obtained at a current variation of 3A and an electroplating time of 30 minutes. The trend of corrosion rate value will decrease with the increase in electric current and the length of electroplating time with the lowest value of 0.003359 mmpy at a variation of electric current 3A and electroplating time of 30 minutes. For optimal values with the lowest corrosion rate on mild steel SS400 JIS G3101 as an oil storage tank material on fuel transport trucks, it is obtained by using a current of 3A and an electroplating time of 30 minutes Ni-Cr.

Improved Artificial Aggregates for Use in Green Roof Design

The development of surfaces in cities, as a result of progressive urbanization, not only reduces the natural retention capacity of the environment but also causes changes in the water balance. In urbanized areas, the amount and intensity of rainwater discharged to receivers increase, and the time of water outflow from the catchment area shortens. Low retention does not provide effective responses to the local water deficit and does not limit the effects of excess water during flood periods. Furthermore, aging drainage systems do not always have the required hydraulic efficiency in absorbing runoff after intense and heavy rainfall or snowmelt. The aim of the work was to determine the possibility of obtaining flat aggregates with a grain size of 2–16 mm from clay-silt fractions from sedimentation tanks using selected mechanical processing methods (crushing and screening in a crusher-screener system with recycling). An important issue was the examination of the physical and mechanical properties of the produced aggregates after firing, where the work required a detailed material analysis using various research techniques, such as XRD, XRF, SEM and digital microscopy. The obtained results will allow for further research on developing the concept of technology for the production of lightweight aggregates used, for example, on building roofs. Particular attention was paid to the flat shape of the aggregate, which affects a number of its properties. To obtain a flat-shaped aggregate, the authors used a patented sieving method. The obtained materials had high cavernosity of 69% on average, water absorption of 40.7% and low bulk density of 0.82 g/cm3.

Correlative Raman Imaging: Development and Cancer Applications.

Despite extensive research efforts, cancer continues to stand as one of the leading causes of death on a global scale. To gain profound insights into the intricate mechanisms underlying cancer onset and progression, it is imperative to possess methodologies that allow the study of cancer cells at the single-cell level, focusing on critical parameters such as cell morphology, metabolism, and molecular characteristics. These insights are essential for effectively discerning between healthy and cancerous cells and comprehending tumoral progression. Recent advancements in microscopy techniques have significantly advanced the study of cancer cells, with Raman microspectroscopy (RM) emerging as a particularly powerful tool. Indeed, RM can provide both biochemical and spatial details at the single-cell level without the need for labels or causing disruptions to cell integrity. Moreover, RM can be correlated with other microscopy techniques, creating a synergy that offers a spectrum of complementary insights into cancer cell morphology and biology. This review aims to explore the correlation between RM and other microscopy techniques such as confocal fluoresce microscopy (CFM), atomic force microscopy (AFM), digital holography microscopy (DHM), and mass spectrometry imaging (MSI). Each of these techniques has their own strengths, providing different perspectives and parameters about cancer cell features. The correlation between information from these various analysis methods is a valuable tool for physicians and researchers, aiding in the comprehension of cancer cell morphology and biology, unraveling mechanisms underlying cancer progression, and facilitating the development of early diagnosis and/or monitoring cancer progression.

Experimental Study on the Flexural Performance of Steel-Polyvinyl Alcohol Hybrid Fiber-Reinforced Concrete.

This paper explores the impact of steel-PVA hybrid fibers (S-PVA HF) on the flexural performance of panel concrete via three-point bending tests. Crack development in the concrete is analyzed through Digital Image Correlation (DIC) and Scanning Electron Microscope (SEM) experiments, unveiling the underlying mechanisms. The evolution of cracks in concrete is quantitatively analyzed based on fractal theory, and a predictive model for flexural strength (PMFS) is established. The results show that the S-PVA HF exhibits a synergistic effect in enhancing and toughening the concrete at multi-scale. The crack area of steel-PVA hybrid fiber concrete (S-PVA HFRC) is linearly correlated with deflection (δ), and it further reduces the crack development rate and crack area compared to steel fiber-reinforced concrete (SFRC). The S-PVA HF improves the proportional ultimate strength (fL) and residual flexural strength (fR,j) of concrete, and the optimal flexural performance of concrete is achieved when the steel fiber dosage is 1.0% and the PVA fiber dosage is 0.2%. The established PMFS of hybrid fiber-reinforced concrete (HFRC) can effectively predict the flexural strength of concrete.

Surface characterization of consolidated earthen substrates using an innovative multi-analytical strategy

This study evaluates the combined use of innovative non-destructive methods and standard laboratory techniques for assessing the surface effects of various consolidation treatments (i.e., bacterial biomineralization, alkaline activation, colloidal dispersions containing nanolime or nanosilica, and ethyl silicate as a standard) on earthen building materials. It demonstrates that novel portable equipment (i.e., portable digital microscope, air permeameter, and mobile surface (contact angle) analyzer) and advanced strategies for the application and data interpretation of conventional analytical and testing methods (i.e., scanning electron and confocal microscopy, Leeb hardness testing, and visible light spectrophotometry) can yield complementary and extremely valuable quantitative results for the evaluation of consolidation treatments on such complex substrates. Our multi-analytical approach allows a detailed characterization of surface and near-surface features of treated earthen mock-ups. It is possible to disclose significant differences in the film-forming propensity and in-depth distributions of consolidants and their effect on key properties such as surface roughness and appearance, permeability, water behavior, and mechanical properties. Even though, the substrate's inhomogeneity and surface texture has proven to be especially challenging, this methodology offers a promising strategy for long-term in-situ monitoring of treatment performance and weathering behavior of built heritage materials.

Effects of Fungicides on Fungal Development, Conidiophore Morphology, and Conidial Releases from Single Strawberry Powdery Mildew Colonies Assessed Using an Electrostatic Technique

We evaluated the effects of fungicides on strawberry powdery mildew (PM) fungus, Podosphaera aphanis, using an electrostatic technique. Thirty-six fungicides were sprayed on single colonies of P. aphanis on leaves of strawberry seedlings (Fragaria × ananassa Duchesne ex Rozier). Colony development varied depending on the tested fungicides. Particularly, pyraziflumid, triflumizole, triforine, polyoxin, sodium hydrogen carbonate + copper wettable powder, and flutianil + mepanipyrim were highly effective for reducing colony development. P. aphanis colonies were histochemically stained to observe the morphological characteristics of fungal cells forming normal and abnormal conidiophores. Abnormal conidiophores were classified into seven types based on their morphological and cytological characteristics. Finally, asexual conidia were collected from single P. aphanis colonies on the leaves spray-treated with fungicides using a dielectrically polarized insulator plate (electrostatic spore collector); conidia attracted to the insulator plates were counted using a high-fidelity digital microscope. Most tested fungicides highly inhibited the production and/or germination of asexual conidia. The germination of asexual conidia was observed only in thiophanate-methyl (methyl benzimidazole carbamates fungicides; MBC fungicides) and azoxystrobin (quinone outside inhibitors; QoI fungicides). Assessing with the electrostatic technique, we clarified that P. aphanis has developed resistance to both thiophanate-methyl and azoxystrobin. Thus, the methodological assessment analyzing the colony development and the number of conidia released from single colonies will be helpful information for screening effective fungicides.

Performance of carbon nanomaterials incorporated with concrete exposed to high temperature

Abstract In recent decades, there have been initiatives to incorporate carbon nanomaterials (CNMs) into cement composites, particularly graphene oxide (GO), carbon nanotubes, graphite (GP), and mild carbon (MC). Nevertheless, little is known about how these CNMs interact with the cement matrix itself. In this research, the impact of CNM incorporation at high temperatures (250, 500, 750, and 1,000°C) on cement’s mechanical characteristics and microstructure was investigated. Nine mixes were created with the CNM content (0.1 and 0.3%) being taken into consideration. The microstructure of the CNM composites was further investigated using X-ray diffractometry, thermogravimetry, derivative thermogravimetry, digital microscopy, and micro-computed tomography (micro-CT). Based on research observations, the study demonstrated that the mechanical properties of most specimens could be enhanced through the introduction of CNMs. The recommended proportions of GP-0.1, GO-0.1, and MC-0.1, in accordance with the weight of the binder, and the impact of the CNMs on the elastic modulus were also assessed. As a consequence, the CNM’s porous structure and apparent crack pattern were identified using microstructure analysis.

Assessment of Indoor and Outdoor Resting Adult Female Mosquitoes Density Exhibiting Vectorial Portent, Insight from Kano State, Nigeria

Mosquito proliferation has been on the increase in most communities of Kano State, leading to a high rate of mosquito-borne diseases, pregnant women, and infant mortality. Mosquitoes spend the daytime resting in human dwellings, frequent survey of these mosquitoes is needed in vector control for breaking disease transmission. Despite this, data on mosquitos’ density are frequently missing. In this study, we have collected and identified all samples of indoor and outdoor resting mosquito species to assess female density. A longitudinal survey was carried out in three different sentinel sites (Darmanawa, Wudil, and Baba Ruga) for the period of 6 months (August 2023 to March 2024). Indoor mosquitoes were collected using the pyrethrum spray Method, while the outdoor mosquitoes were collected using a cardboard box. The knockdown mosquitoes were transported to the laboratory for identification, using a digital LCD microscope to gender and species level with the aid of different taxonomic guides. A total of one thousand and five (1005) female mosquitoes belonging to 5 genera and 21 species were caught. The result showed a significant influence (p &lt; 0.05) of species, months, resting behavior, and season on the density of adult female mosquitoes while collection sites do not influence adult female mosquitoes (p &gt; 0.05). We concluded that adult female mosquitos’ density is affected by species, months of the year, resting behavior, and season, as well as the high density of An. gambiae and Cx. quinnquefasciatus recorded is a bio-portent of malaria and filariasis spotlight in these communities.

Transformer based deep learning hybrid architecture for phase unwrapping

A deep learning Hybrid architecture for phase unwrapping has been proposed. The hybrid architecture is based on integration of Convolutional Neural Networks (CNN) with Vision Transformer. The performance of Hybrid architecture/network in phase unwrapping is compared against CNN based standard UNET network. Structural Similarity Index (SSIM) and Root Mean Square Error (RMSE) have been used as performance metrics to assess the performance of these deep learning networks for phase unwrapping. To train and test the networks, dataset with high mean Entropy has been generated using Gaussian filtering of random noise in Fourier plane. The Hybrid architecture is tested on test dataset and is found to have superior performance metrics against the UNET network. Their performance is also tested in noisy environment with various noise levels and Hybrid architecture demonstrated better anti-noise capability than UNET network. Hybrid architecture was successfully validated in real world scenario using experimental data from custom built Digital Holographic Microscope. With the advent of newer architectures and hardware, Deep learning networks can further improve the performance in solving inverse problems.

Direct GPCR-EGFR interaction enables synergistic membrane-to-nucleus information transfer

We addressed the heteromerization of the epidermal growth factor receptor (EGFR) with G-protein coupled receptors (GPCR) on the basis of angiotensin-II-receptor-subtype-1(AT1R)-EGFR interaction as proof-of-concept and show its functional relevance during synergistic nuclear information transfer, beyond ligand-dependent EGFR transactivation. Following in silico modelling, we generated EGFR-interaction deficient AT1R-mutants and compared them to AT1R-wildtype. Receptor interaction was assessed by co-immunoprecipitation (CoIP), Förster resonance energy transfer (FRET) and fluorescence-lifetime imaging microscopy (FLIM). Changes in cell morphology, ERK1/2-phosphorylation (ppERK1/2), serum response factor (SRF)-activation and cFOS protein expression were determined by digital high content microscopy at the single cell level. FRET, FLIM and CoIP confirmed the physical interaction of AT1R-wildtype with EGFR that was strongly reduced for the AT1R-mutants. Responsiveness of cells transfected with AT1R-WT or –mutants to angiotensin II or EGF was similar regarding changes in cell circularity, ppERK1/2 (direct and by ligand-dependent EGFR-transactivation), cFOS-expression and SRF-activity. By contrast, the EGFR-AT1R-synergism regarding these parameters was completely absent for in the interaction-deficient AT1R mutants. The results show that AT1R-EGFR heteromerisation enables AT1R-EGFR-synergism on downstream gene expression regulation, modulating the intensity and the temporal pattern of nuclear AT1R/EGFR-information transfer. Furthermore, remote EGFR transactivation, via ligand release or cytosolic tyrosine kinases, is not sufficient for the complete synergistic control of gene expression.

Hematological cytomorphology: Where we are.

The manuscript discusses the historical evolution of observing blood cell morphology under an optical microscope, from the earliest microscopes in the 17th century to the modern digital era, highlighting key advancements and contributions in the field. Blood has historically held symbolic importance in various cultures, with early medical observations dating back to Hippocrates and Galeno. The discovery of cells and subsequent advancements in microscopy by scientists like Hooke and van Leeuwenhoek paved the way for understanding blood cell morphology. Influential figures such as Hewson, Donné, and Ehrlich followed. Diagnostic cytology evolved from manual cell counting to the development of automated hematological systems. Automated complete blood counting came to support microscopic examination in diagnosing hematological disorders. Morphology is crucial in predicting disease outcomes and guiding treatment decisions, particularly hematological neoplasms. The introduction of flow cytometry and its integration with traditional morphological analysis and the new cytogenetic and molecular techniques revolutionized the classification and prognostication of hematologic disorders. Digital microscopy has emerged as a powerful tool in recent years, offering rapid acquisition and sharing of blood cell images. Integrating Artificial Intelligence with digital microscopy has further enhanced morphological analysis, improving diagnostic efficiency. We also discuss the prospects of AI in pre-classifying blood cells in bone marrow aspirate samples, potentially revolutionizing diagnostic pathways for hematologic diseases. Overall, the manuscript provides a comprehensive overview of the historical development, clinical significance and technological advancements in observing blood cell morphology, underscoring its continued relevance in modern hematology practice.