Bright Field Research Articles

Dual‐Functional X‐Ray Photochromic Phosphor: High‐Performance Detection and 3D Imaging

AbstractThis study explores the integration of X‐ray‐induced photochromism and photoluminescence in a single material, offering innovative multi‐mode applications in optical memory, anti‐counterfeiting, and X‐ray detection and imaging. The photochromic phosphor LiAlSi2O6:Sm3+ is synthesized, which undergoes a color change from white to dark green under bright field conditions and displays orange photoluminescence in dark field conditions after X‐ray exposure due to defect formation. The phosphor shows accelerated bleaching and recovery under 473 nm laser stimulation. The distinct X‐ray‐induced color contrast and luminescence intensity modification in LiAlSi2O6:Sm3+ highlight its potential in advanced luminescent material design. Moreover, a LiAlSi2O6:Sm3+‐based flexible film demonstrates “dual‐mode” 3D X‐ray imaging and detection capabilities, paving the way for future X‐ray detecting and imaging device research.

Effect of mango butter on the physicochemical properties of beeswax-Moringa seed oil-based oleogels for topical application

Objective The purpose of this research was to determine any connections between the characteristics of oleogels made of beeswax and the impact of mango butter. Methods Oleogel was prepared through inverted tube methods, and optimized through oil binding capacity. Other evaluations like bright field and polarized microscopy, Fourier-transform infrared (FTIR) spectroscopy, crystallization kinetics, mechanical study, and X-ray diffractometry (XRD). The drug release kinetic studies and in vitro antibacterial studies were performed. Results FTIR study reveals that the gelation process does not significantly alter the chemical composition of the individual components. Prepared gel exhibiting fluid-like behavior or composed of brittle networks is particularly vulnerable to disruptions in their network design. The incorporation of mango butter increases the drug permeation. In-vitro microbial efficacy study was found to be excellent. Conclusion The studies revealed that mango butter can be used to modify the physico-chemical properties of the oleogels.

Multimodal Spectroscopy Assays for Advanced Nano-Optics Approaches by Tuning Nano-Tool Surface Chemistry and Metal-Enhanced Fluorescence

In this research work, different chemical modifications were applied to gold nanoparticles and their use in enhanced non-classical light emitters based on metal-enhanced fluorescence (MEF) was evaluated. In order to achieve this, gold core–shell nanoparticles with silica shells were modified via multilayered addition and the incorporation of a covalently linked laser dye to develop MEF. Their inter-nanoparticle interactions were evaluated by using additional silica shell multilayers and modified cyclodextrin macrocycles. In this manner, the sizes and chemical surface interactions on the multilayered nanoarchitectures were varied. These optical active nanoplatforms led to the development of different nanoassembly sizes and luminescence behaviors. Therefore, the interactions and nanoassembly properties were evaluated by using various spectroscopic and nanoimaging techniques. Highly dispersible gold core–shell nanoparticles with diameters of 50–60 nm showed improved colloidal dispersion that led to single ultraluminescent gold core–shell nanoparticles with MEF. Then, the addition of variable silica lengths produced increased interactions and consequent nanoaggregation. However, the silanized nanoparticles were easily dispersible after agitation or sonication. Thus, their sizes were proportional only to the diameter and the van de Waals interaction did not affect their sizes in bulk. Then, the covalent linking of different concentrations of modified cyclodextrins was applied to the chemical surfaces by incorporating additional hydroxyl groups from the glucose monomeric unities of cyclodextrins. In this manner, variable larger-sized and inter-branched grafted gold core–shell silica nanoparticles were generated. The ultraluminescent properties were conserved due to the non-optical activity of the cyclodextrins. However, they generated enhanced ultraluminescence phenomena. Laser fluorescence microscopy nanoimaging showed enhanced resolutions in comparison to non-grafted supramolecular gold core–shell nanoparticles. The differences in their interactions and the sizes of the nanoassemblies were explained by their single nanoparticle diameters and the interacting chemical groups on their nanosurfaces. While the varied luminescence emissions generated were tuned by plasmonics, enhanced plasmonic phenomena and light scattering properties were seen depending on the type of nanoassembly. Thus, optically active and non-optically active materials led to different optical properties in the bright field and enhanced the excited state within the electromagnetic near-field of the gold nanotemplates. In this manner, it was possible to achieve high sensitivity by varying the spacer lengths and optical properties. Therefore, further perspectives regarding the design of nano-tools composed of light for various applications were discussed.

Image-based profiling and deep learning reveal morphological heterogeneity of colorectal cancer organoids

Patient-derived organoids have proven to be a highly relevant model for evaluating of disease mechanisms and drug efficacies, as they closely recapitulate in vivo physiology. Colorectal cancer organoids, specifically, exhibit a diverse range of morphologies, which have been analyzed with image-based profiling. However, the relationship between morphological subtypes and functional parameters of the organoids remains underexplored. Here, we identified two distinct morphological subtypes (“cystic” and “solid”) across 31360 bright field images using image-based profiling, which correlated differently with viability and apoptosis level of colorectal cancer organoids. Leveraging object detection neural networks, we were able to categorize single organoids achieving higher viability scores as “cystic” than “solid” subtype. Furthermore, a deep generative model was proposed to predict apoptosis intensity based on a apoptosis-featured dataset encompassing over 17000 bright field and matched fluorescent images. Notably, a significant correlation of 0.91 between the predicted value and ground truth was achived, underscoring the feasibility of this generative model as a potential means for assessing organoid functional parameters. The underlying cellular heterogeneity of the organoids, i.e., conserved colonic cell types and rare immune components, was also verified with scRNA sequencing, implying a compromised tumor microenvironment. Additionally, the “cystic” subtype was identified as a relapse phenotype featuring intestinal stem cell signatures, suggesting that this visually discernible relapse phenotype shows potential as a novel biomarker for colorectal cancer diagnosis and prognosis. In summary, our findings demonstrate that the morphological heterogeneity of colorectal cancer organoids explicitly recapitulate the association of phenotypic features and exogenous perturbations through the image-based profiling, providing new insights into disease mechanisms.

Survey of Southeast U.S. beef veterinarians regarding methods for bull breeding soundness evaluation

The purpose of this survey was to describe veterinarians’ methods for performing bull breeding soundness evaluations (BSEs). Veterinarians in Mississippi, Louisiana and Arkansas were emailed a survey regarding their experience, demo­graphics, opinions and perceptions of client pref­erences regarding BSEs. Logistic regression was used to test respondent characteristics for associations with dichotomous outcomes including evaluating semen morphology as part of every BSE and detecting the diadem defect. Of 3,757 veterinar­ians solicited, 204 (5%) responded and 83 (2%) qualified for analysis. Of these, 10/73 (14%) indicated they do not evaluate morphology during every BSE. When shown an image of the diadem defect, 18/73 (25%) indicated seeing it often or some­times, and 55/73 (75%) indicated rarely or never. The only fac­tor associated with evaluating morphology as part of every BSE, was indicating belief that morphology was most predic­tive of bull fertility vs. believing motility was most predictive, or being unsure (OR = 11.2, 95% C.I. = 1.3-94.1). Respondents who always evaluate morphology with bright field microscopy at 1,000X (OR = 4.1, 95% C.I. = 1.2-13.3) or with phase contrast microscopy at ≥ 400X (OR = 5.4, 95% C.I. = 1.1-27.7) had higher odds of indicat­ing they detect the diadem defect sometimes/often. There was an interaction between always using 1000X magnification, or not, and always using 400X, or not, on the reported detection of the diadem defect. Veterinarians’ perceptions of which as­pects of the BSE are most predictive of fertility influence their methods for performing BSEs, and influence the frequency of detecting diadem defects.

Abstract 700: Impact of heterogeneity for mismatch repair activity on colon tumor development and therapeutic response

Abstract Background: Colorectal cancers (CRC) that are deficient for DNA mismatch repair (dMMR) proteins have high levels of genetic instability and consequently high mutational burden, creating numerous neoantigens that can elicit an immune response. Therefore, dMMR CRC are the best candidates for immune checkpoint inhibition (ICI) compared to proficient DNA mismatch repair (pMMR) CRC. Early clinical trials revealed that only 30-55% metastatic dMMR CRC responded to ICI. Recent studies utilizing allograft models have demonstrated that tumor immune landscape can be primed with radiopharmaceutical therapy (RPT). Although early results have been quite promising, allograft models often fail to accurately predict efficacy for many reasons, but most often are due to a lack of heterogeneity. Methods: To better understand how heterogenous MMR expression impacts tumor development and response to ICI alone or in combination with RPT, we developed a new mouse model that develops three tumor types: (1) homotypic dMMR tumors that express green fluorescent protein, (2) homotypic pMMR tumors that express red fluorescent protein, and (3) heterotypic tumors with a mixture of dMMR and pMMR cells. Surveillance bright field and fluorescent colonoscopies allow us to assess treatment response by measuring tumor size and proportion of dMMR and pMMR tumor cells in real-time. These outcomes are verified through ex vivo imaging and histological analysis. Results: Tumor response to anti-PD-L1 ICI was dependent on MMR status. Homotypic dMMR tumors exhibited the strongest response: significantly smaller in treated mice than controls, exhibited areas of vascular congestion and increased CD8+ cytotoxic T-cells infiltration. Response of heterotypic MMR tumors varied depending on percentage of dMMR cells; tumors with a high percentage of dMMR cells remained small, whereas tumors with a low percentage of dMMR had an outgrowth of pMMR cells. Treated and untreated homotypic pMMR tumors were indistinguishable. To enhance the response of all three tumor types, mice were treated with dual ICI treatment, anti-PD-L1 and anti-CTLA-4, or RPT. Animals administered the tri-therapy (dual ICI + RPT) survived significantly longer with fewer tumors than animals treated with either dual ICI or RPT alone. Conclusions: Our innovative mouse model allows for the highly detailed characterization of tumor response to therapies. Simplistic allograft models may not adequately account for the dynamic effects that neighboring cell death may have on radio-resistant cells, on tumor vascular perfusion and oxygenation, or on tumor immune infiltration and adaptive immune recognition of remaining cells. We begin to fill these critical gaps in knowledge and therefore the results from our studies will help guide the translation and integrated use of ICIs and RPTs in clinic. Citation Format: Santina M. Snow, Dawn Albrecht, Paul A. Clark, Caroline P. Kerr, Joseph J. Grudzinski, Justin J. Jeffery, Hansel Comas Rojas, Reinier Hernandez, Kristina A. Matkowskyj, Jamey P. Weichert, Zachary S. Morris, Richard B. Halberg. Impact of heterogeneity for mismatch repair activity on colon tumor development and therapeutic response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 700.

Abstract 221: Novel organotypic patient derived primary tumor tissues for oncology drug safety and efficacy studies

Abstract Organotypic, three-dimensional (3D) cancer tissue models (OCTM) have enabled investigators to develop predictive markers that can be used in preclinical cancer drug development. However, a drawback of the existing models is that they are mostly based on cell lines based are not physiological. In vitro development of 3D tumoroids using patient derived primary tumors (PDPT) in tumor microenvironment (TME) will have clinical significant and can predict human responses to cancer therapeutics. Here, we describe the generation of OCTM from patient-derived metastatic colorectal cancer, the second most deadly cancer. PDPT were grown in Matrigel for 5 passages and for biobanking purposes tumoroids were further expanded in faCellitate 384 well plates or Sun Bioscience plates to harvest large amounts of well-defined tumoroids cultured for over 21 days. Expanded tumoroids were collected and cryopreserved or gently mixed several times using a pipet tip to break up tumoroids into smaller fragments. These tumor fragments were mixed with endothelial cells, dendritic cells, fibroblasts and embedded in a collagen matrix to simulate TME. The collagen-cell mixture was then added to Transwells and induced to gel to form a collagen gel matrix that simulates TME. The tumoroids were cultured with a specialized medium that supports the growth of complex cell types. The tumor development was monitored daily using bright field microscopy and image analysis was used to monitor tumor size. To evaluate the utility of the OCTM, treatment was initiated at day 10 of the culture period by exposing tissues with the chemotherapeutic drug Cisplatin on days 0, 2, 4, 7, 9, and 11. After each treatment, OCTM tissues (N=2) were fixed for H&E and live/dead staining. Overall, the results of the study showed that: 1) PDPT can be expanded for 5 passages (this low passage number is not expected to change the phenotype of the tumor) and further expanded in high throughput format, 384 well plates, 2) the PDPT could be cultured with multiple cell types in a collagen-gel matrix to recreate the TME, 3) histological evaluation of the OCTM showed a glandular-like tumoroid, 4) immunohistochemical staining revealed that the PDPT in the in vivo-like microenvironment were positive for CK20, and 5) the PDPT shrank following 3 treatment cycles with Cisplatin (day 4). Conclusion: The data shows metastatic colorectal PDPT can be expanded in vitro and cultured in a complex tumor microenvironment on tissue culture inserts. These OCTMs can be used to screen drug safety and efficacy. This study opens new avenues for high-throughput cancer drug screening format using PDPT embedded in a in vivo like TME. This model can be used as a tool for personalized oncology drug testing and for translational assays tailored to other cancer types from our patient derived primary tumor pool. Citation Format: Seyoum Ayehunie, Megan Groves, Bryda Bryda, Alex Armento, Anthony Tolcher. Novel organotypic patient derived primary tumor tissues for oncology drug safety and efficacy studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 221.

Abstract 2103: Small extracellular vesicles derived from macrophages show anti-tumor effects in MDA-MB-231 cells

Abstract Triple-negative breast cancer is the primary cause of death in women globally due to the absence of targeted receptors on the tumor, making it hard to treat. In addition, the rapid spread of triple-negative breast cancer makes it difficult to manage using current treatment options such as lumpectomies, mastectomies, radiation, and chemotherapy. The survival rate for patients with triple-negative breast cancer is only 8-16%. Therefore, there is a critical need to develop new and effective treatment options. The objective of this study was to investigate the potential of M1 macrophage-derived exosomes to induce cancer cell apoptosis. To test this, we isolated M1 macrophage-derived exosomes to see their effects on triple-negative breast cancer MDA-MB-231 cells. We induce polarization in RAW 264.7 macrophage cells via lipopolysaccharide (LPS). Macrophage polarization was characterized using an enzyme-linked immunosorbent assay (ELISA), real-time PCR, nitric oxide production, and bright field microscopy. We characterized exosomes using nanoparticle tracking analysis (NTA) and scanning electron microscopy (SEM). The apoptosis marker, caspase 3/7 activation, was assessed using confocal microscopy in MDA-MB-231 cells. M1 macrophage-derived exosomes were isolated, characterized, and incubated with breast cancer cells to see their anticancer effects. Data showed the induction of apoptosis in 48 hours in triple-negative breast cancer cells. We conclude that exosomes derived from M1 macrophages may have the capability to induce apoptosis in triple-negative breast cancer. Citation Format: Parth Desai, Anjali Kumari, Saqer Al Abdullah, Kristen Dellinger. Small extracellular vesicles derived from macrophages show anti-tumor effects in MDA-MB-231 cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2103.

Abstract 6185: Generative AI for virtual HE-staining of whole slide images of unstained breast cancer tissue

Abstract Introduction Hematoxylin and eosin (HE) is the standard stain used in histology to make tissue visible to the human eye by highlighting certain cellular and tissue structures, and it is a technique that is widely used in the diagnosis of cancer and other pathologies. Chemical staining, however, is irreversible, making the tissue unusable for subsequent measurements, such as spatial transcriptomics. Here we utilize generative AI method based on pix2pix image-to-image translation to generate virtual HE-staining for whole slide images (WSIs) acquired of unstained tissue with brightfield microscopy and perform a thorough histological evaluation for the feasibility in breast cancer diagnostics. Materials and Methods We optimized sample preparation and imaging setup for virtual staining purposes, and developed a custom generative adversarial network architecture for learning the virtual staining from paires samples of unstained and H&E stained tissue. Here, we focus on the utility of virtual staining in breast cancer diagnostics, using a set of breast cancer samples for acquiring whole slide images from unstained tissue before H&E staining and from reference H&E stained tissue after chemical staining. Hold-out set of sample pairs are left for validation, allowing us to evaluate the virtual staining performance for a vast array of tissue components and to examine the potential shortcomings in staining reproduction. We use a comprehensive set of quantitative metrics both on pixel and object level to evaluate virtual staining quality. In addition, we perform thorough visual evaluation of histological feasibility by histology experts to examine the computational staining. Results We demonstrate that by careful optimization of both sample preparation and imaging workflow, as well as the computational methods, generative adversarial networks can be used for virtual staining of whole slide images of breast cancer tissue acquired from unstained tissue using regular bright field microscopy. We analyzed the virtual staining performance quantitatively and visually, and highlight the potential of the method through successful cases and demonstrate the applicability in breast cancer diagnostics, as well as discuss the challenges and shortcomings of virtual staining for clinical samples. Conclusions We demonstrate the potential of virtual HE staining for clinical histopathology of breast cancer tissue. Notably, our virtual staining based on generative AI shows promise towards more sustainable and streamlined sample processing and staining process in digital pathology. Citation Format: Leena Latonen, Umair Khan, Sonja Koivukoski, Johan Hartman, Pekka Ruusuvuori. Generative AI for virtual HE-staining of whole slide images of unstained breast cancer tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6185.

Adipose Tissue and Bone Marrow-Derived Mesenchymal Stem Cells are not Really the Same: Investigating the Differences in Their Immunomodulatory, Migratory, and Adhesive Profile.

Mesenchymal stem cells (MSCs) are the most widely used stem cells in regenerative medicine. They can be isolated from multiple sources, most commonly bone marrow and adipose tissue. MSCs derived from different sources show similar molecular and biological characteristics, but there is ongoing debate regarding the best source of MSCs and the potential biological differences between MSCs from different origins. Bone marrow derived-MSCs (BM-MSCs) and adipose tissue-derived MSCs (AD-MSCs) share many molecular and immunomodulatory properties. In this study, we compared the levels of major immunomodulatory, adhesive, and migratory factors in human BM-MSCs and AD-MSCs under normal conditions, which will help determine the suitability and specificity of each type for certain therapeutic applications. WST1 assay and fluorescent assay SUC-LLVY-AMC were used to measure MSC proliferation and 26S proteasome activity, respectively. Western blotting, ELISA Assays, and bright field live imaging were also used. AD-MSCs and BM-MSCs exhibited similar morphology and proliferation rate. A significantly higher 26S proteasome activity was detected in AD-MSCs than in BM-MSCs. Levels of ICAM-1, integrin α5 and integrin α6 were significantly higher in AD-MSCs compared to BM-MSCs, while no significant difference in CXCR4 levels was observed. Expression of IDO and factor H was significantly higher in AD-MSCs, while CTLA-4 and IL-10 levels were higher in BM-MSCs. This indicates that AD-MSCs and BM-MSCs have different immunomodulatory and adhesion profiles. MSCs isolated from different sources may show differences in their biological and immunomodulatory properties, suggesting a potential suitability of certain MSCs type for specific conditions. Also, combination of different MSCs types could help optimize therapeutic outcomes.

Ambient particulate matter monitoring using bright field imaging-based sensor

The bright-field imaging of the particulate matter is demonstrated with the customized bright-field (BF) microscopic unit. Polystyrene spheres of size 4.5 µm as reference objects have been imaged to validate the claim. An image-processing-based algorithm has been used which uses pixel size for particle sizing and distribution. The lateral resolution of the device is found to be approximately 0.5 µm and the field of view is 56 µm × 42 µm which has been utilized for the imaging of the ambient PM. The device gives the results in the form of particulate matter concentration, size distribution, and shape of particulate matter.

The impact of ionization morphology and X-ray heating on the cosmological 21cm skew spectrum

Abstract The cosmological 21cm signal offers a potential probe of the early Universe and the first ionizing sources. Current experiments probe the spatially-dependent variance (Gaussianity) of the signal through the power spectrum (PS). The signal however is expected to be highly non-Gaussian due to the complex topology of reionization and X-ray heating. We investigate the non-Gaussianities of X-ray heating and reionization, by calculating the skew spectrum (SS) of the 21cm signal using meraxes, which couples a semi-analytic galaxy population with semi-numerical reionization simulations. The SS is the cross-spectrum of the quadratic temperature brightness field with itself. We generate a set of seven simulations from z = 30 to z = 5, varying the halo mass threshold for hosting star-formation, the X-ray luminosity per star-formation rate, and the minimum X-ray energy escaping host galaxies. We find the SS is predominantly negative as a function of redshift, transitioning to positive towards the start of reionization, and peaking during the midpoint of reionization. We do not see a negative dip in the SS during reionization, likely due to the specifics of modelling ionization sources. We normalise the SS by the PS during reionization isolating the non-Gaussianities. We find a trough (k ∼ 0.1 Mpc−1) and peak (k ∼ 0.4 − 1 Mpc−1) in the normalised SS during the mid to late periods of reionization. These correlate to the ionization topology, and neutral islands in the IGM. We calculate the cosmic variance of the normalised SS, and find these features are detectable in the absence of foregrounds with the SKA_LOW.

Convolutional neural network approach for the automated identification of in cellulo crystals.

In cellulo crystallization is a rare event in nature. Recent advances that have made use of heterologous overexpression can promote the intracellular formation of protein crystals, but new tools are required to detect and characterize these targets in the complex cell environment. The present work makes use of Mask R-CNN, a convolutional neural network (CNN)-based instance segmentation method, for the identification of either single or multi-shaped crystals growing in living insect cells, using conventional bright field images. The algorithm can be rapidly adapted to recognize different targets, with the aim of extracting relevant information to support a semi-automated screening pipeline, in order to aid the development of the intracellular protein crystallization approach.

Pernikahan Adat Pepadun Perspektif Pendidikan Islam

This research explores the implementation of the marriage customs of the Lampung Pepadun community in the Sungkai Bunga Mayang clan from an Islamic education perspective. The research background reflects the complexity of Lampung traditional marriages, where traditional traditions and Islamic norms often blend. The traditional Lampung wedding ceremony in the Sungkai Bunga Mayang clan involves Rasan Sanak (elopement) and Rasan Tuho (parental consent). This research aims to analyze and describe the implementation of wedding customs, with a focus on compliance with the principles of Islamic education. The method used is descriptive qualitative research, describing the research object and analyzing it. This research is descriptive in nature which is analyzed qualitatively in order to describe or describe the condition of an object and then analyze it. This research was structured based on field research by collecting data through interviews, observation, documentation and from various literature related to this research study. The results of the research show that the Lampung Pepadun community in the Sungkai Bunga Mayang clan consistently maintains and maintains traditional traditions, such as intar payu, intar bright field, intar bright, intar manum, and sembambangan. This tradition is in line with Islamic education, providing teachings about attitudes and behavior in accordance with Islamic values. However, several marriage practices were found that were not fully in accordance with Islamic teachings, such as high dowries or exchange of goods that were not according to Islamic law. The conclusion of this research highlights the need to increase public awareness of the principles of Islamic education in carrying out traditional Lampung weddings. Efforts to integrate Islamic values ​​into wedding traditions can produce marriages that are more balanced and in accordance with religious teachings.

A Robust Method for the Unsupervised Scoring of Immunohistochemical Staining.

Immunohistochemistry is a powerful technique that is widely used in biomedical research and clinics; it allows one to determine the expression levels of some proteins of interest in tissue samples using color intensity due to the expression of biomarkers with specific antibodies. As such, immunohistochemical images are complex and their features are difficult to quantify. Recently, we proposed a novel method, including a first separation stage based on non-negative matrix factorization (NMF), that achieved good results. However, this method was highly dependent on the parameters that control sparseness and non-negativity, as well as on algorithm initialization. Furthermore, the previously proposed method required a reference image as a starting point for the NMF algorithm. In the present work, we propose a new, simpler and more robust method for the automated, unsupervised scoring of immunohistochemical images based on bright field. Our work is focused on images from tumor tissues marked with blue (nuclei) and brown (protein of interest) stains. The new proposed method represents a simpler approach that, on the one hand, avoids the use of NMF in the separation stage and, on the other hand, circumvents the need for a control image. This new approach determines the subspace spanned by the two colors of interest using principal component analysis (PCA) with dimension reduction. This subspace is a two-dimensional space, allowing for color vector determination by considering the point density peaks. A new scoring stage is also developed in our method that, again, avoids reference images, making the procedure more robust and less dependent on parameters. Semi-quantitative image scoring experiments using five categories exhibit promising and consistent results when compared to manual scoring carried out by experts.

Simultaneous secondary electron microscopy in the scanning transmission electron microscope with applications for in situ studies.

Scanning/transmission electron microscopy (STEM) is a powerful characterization tool for a wide range of materials. Over the years, STEMs have been extensively used for in situ studies of structural evolution and dynamic processes. A limited number of STEM instruments are equipped with a secondary electron (SE) detector in addition to the conventional transmitted electron detectors, i.e. the bright-field (BF) and annular dark-field (ADF) detectors. Such instruments are capable of simultaneous BF-STEM, ADF-STEM and SE-STEM imaging. These methods can reveal the 'bulk' information from BF and ADF signals and the surface information from SE signals for materials <200 nm thick. This review first summarizes the field of in situ STEM research, followed by the generation of SE signals, SE-STEM instrumentation and applications of SE-STEM analysis. Combining with various in situ heating, gas reaction and mechanical testing stages based on microelectromechanical systems (MEMS), we show that simultaneous SE-STEM imaging has found applications in studying the dynamics and transient phenomena of surface reconstructions, exsolution of catalysts, lunar and planetary materials and mechanical properties of 2D thin films. Finally, we provide an outlook on the potential advancements in SE-STEM from the perspective of sample-related factors, instrument-related factors and data acquisition and processing.

Enhancement of Entanglement via Incoherent Collisions.

In contrast to the general thought that the collisions are intrinsically dephasing in nature and detrimental to quantum entanglement at room or higher temperatures, here, we show that in the conventional ladder-type electromagnetically induced transparency (EIT) configuration, when the probe field intensity is not very weak as compared to the pump field, the entanglement between the bright pump and probe fields can be remarkably enhanced with the increase of the collisional decay rates in a moderate range in an inhomogeneously broadened atomic system. The strengthened entanglement results from the enhancement of constructive interference and suppression of destructive interference between one-photon and multiphoton transition pathways. Our results clearly indicate that the collisions offer a promising alternative to enhance entanglement at room or higher temperatures despite of the dephasing nature, which provides great convenience for experimental implementation, and opens new prospects and applications in realistic quantum computation and quantum information processing.

Wedding Customs of the Lampung Pepadun Community in the Perspective of Islamic Education

Lampung traditional marriage ceremony, in its implementation, the marriage of the Sungkai Bunga Mayang clan community is carried out by Rasan Sanak or Sembambangan (elopement) and Rasan Tuho or Intar Sapon (parental approval). Lampung traditional wedding traditions are still many people who carry out wedding customs that do not comply with applicable customary regulations. In some Lampung wedding customs, there may be certain practices such as very high dowries, exchange of goods that are not in accordance with Islamic law, or certain traditions. who do not have basic Islamic education. This research aims to: Analyze and describe the implementation of the marriage customs of the Lampung Pepadun community in the Sungkai Bunga Mayang clan from an Islamic education perspective. This research is descriptive in nature which is analyzed qualitatively in order to describe or describe the condition of an object and then analyze it. This research was structured based on field research by collecting data through interviews, observation, documentation and from various literature related to this research study. In carrying out weddings, the people of Lampung Pepadun in the Sungkai Bunga Mayang clan community always preserve and maintain the customs of intar payu, intar bright field, intar bright, intar manum and sebambangan. This form of tradition is in accordance with Islamic education, this is because the marriage customs of the Lampung Pepadun people in the Sungkai Bunga Mayang clan community are the formation of a Muslim personality, both containing teachings about personal or community attitudes and behavior, towards the welfare of individual and collective life, in accordance with Islamic education. which is individual education and community education.&#x0D; Keywords: Wedding Customs of Lampung, Islamic Education, Lampung Pepadun Community

Demonstration and STEM Analysis of Ferroelectric Switching in MOCVD‐Grown Single Crystalline Al0.85Sc0.15N

AbstractWurtzite‐type Al1−xScxN solid solutions grown by metal organic chemical vapor deposition are for the first time confirmed to be ferroelectric. The film with 230 nm thickness and x = 0.15 exhibits a coercive field of 5.5 MV cm−1 at a measurement frequency of 1.5 kHz. The single crystal quality and homogeneous chemical composition of the film are confirmed by X‐ray diffraction and spectroscopic methods such as time of flight secondary ion mass spectrometry. Annular bright field scanning transmission electron microscopy serves to prove the ferroelectric polarization inversion at the unit cell level. The single crystal quality further allows to image the large‐scale domain pattern of a wurtzite‐type ferroelectric for the first time, revealing a predominantly cone‐like domain shape along the c‐axis of the material. As in previous work, this again implies the presence of strong polarization discontinuities along this crystallographic axis, which can be suitable for current transport. The domains are separated by narrow domain walls, for which an upper thickness limit of 3 nm is deduced but which can potentially be atomically sharp. The authors are confident that these results will advance the commencement of the integration of wurtzite‐type ferroelectrics to GaN as well as generally III‐N‐based heterostructures and devices.

Intact matrix-assisted Golgi expansion visualized by bright and dark field microscopy

Intact matrix-assisted Golgi expansion visualized by bright and dark field microscopy