Electron Microscopy Laboratory Research Articles

Correlation of light and electron microscopic morphometric parameters of glomerular capillaries with serum creatinine and proteinuria

ABSTRACT Waste products in the bloodstream are filtered by the glomerular capillaries in the kidneys and excreted into the urine. When making a differential diagnosis of kidney diseases, structural assessment of glomeruli using histological, ultrastructural, and immunological studies is crucial. This study assessed the microscopic and ultrastructural morphometric parameters of glomerular capillaries and examined their correlation with serum creatinine and proteinuria. A total of 60 kidney biopsy cases received by the transmission electron microscope (TEM) laboratory for diagnosis were included in the study. Toluidine blue stained 300 nm thick sections of TEM tissue blocks were scanned for glomerular morphometry by a whole slide imaging system, and the estimation of Bowman’s capsule (BC) area, glomerular capillary lumen diameter (GCLD), glomerular capillary density (GCD), glomerular capillary surface area density (GCSA), and percentage of glomerular capillary lumen space (%GCLS) was performed with QuPath software. TEM images of 70 nm thick sections were used for the evaluation of endothelial fenestration diameter (EFD), glomerular basement membrane (GBM) thickness, and podocyte foot process (PFP) effacement. Proteinuria and serum creatinine showed positive correlations with GBM thickness and PFP effacement. Negative correlations of serum creatinine were observed with EFD, %GCLS, and GCSA. Hence, glomerular filtration is greatly affected by the total area of the glomerular capillary surface and structural changes of GBM. Reduction of glomerulus filtration due to foot process effacement and thickening of GBM results in damage to the filtration barrier leading to the leakage of plasma protein into urine.

Creating Efficient Workflows for Electron Microscopy Laboratories with Automated Specimen Preparation

Abstract Electron microscopy (EM) is essential to the biological and biomedical sciences and clinical diagnostic pathology. Electron microscopy specimen preparation is laborious and time-consuming, with transmission EM (TEM) chemical preparation typically requiring 1–3 days, while volume electron microscopy (vEM) takes 3–5 days of tedious manual reagent exchanges every few minutes or hours. This places a considerable burden on laboratory scientists with the ongoing demand for TEM, and the rapidly growing demand for vEM due to its potential to revolutionize structural biology, connectomics, and related fields. This burden is exacerbated by a shortage of trained electron microscopy scientists as current staff retire, and few enter the workforce. This report provides four case studies to illustrate how automated and faster specimen preparation workflows using mPrep™ Automated Specimen Processors (ASP-1000™ and ASP-2000™, Microscopy Innovations, LLC) free electron microscopy staff in academic and pre-clinical research labs and a clinical pathology laboratory.

Indigenous artefacts and oral stories

This article presents the results of studies conducted by Canadian academics in collaboration with Sturgeon Lake and Pelican Narrows First Nations communities (Saskatchewan, Canada). The objectives of the project were: (1) developing a research ethics protocol for collecting, studying, and preserving indigenous artefacts; (2) measurements of chemical compositions of artefacts; (3) collecting oral stories of Elders. Within the project, two workshops were organized in Pelican Narrows and Sturgeon Lake. Post-secondary students were trained to work on the project. The laboratory measurements of chemical compositions of artefacts were conducted at the Scanning Electron Microscope Laboratory (University of Alberta), and the Saskatchewan Isotope Laboratory (University of Saskatchewan). The carbon-dating measurements were carried out at André E. Lalonde Accelerator Mass Spectrometry Laboratory (University of Ottawa). The statistical analysis of chemical compositions was conducted in order to test the provenance similarities of artefacts. The project was supported by the Department of Canadian Heritage.

Studies of Physical Parameters of Indigenous Artifacts. Collecting and Preserving the Relating Oral Stories

The project, supported by the Department of Canadian Heritage, was conducted by scholars from First Nations University of Canada (FNUniv), University of Regina (U of R) and Royal Saskatchewan Museum (RSM) in 2018 - 2020, in collaboration with Sturgeon Lake and Pelican Narrows First Nations communities. More than forty people, including Elders and students, participated in this project. The research Ethics review was done by U of R. We also consulted with Elders regarding the research Ethics protocols during the individual meetings, ceremonies and workshops in Pelican Narrows and Sturgeon Lake. Research assistants (Indigenous students) were trained (Indigenous studies and the basics of archaeology) for working in First Nations communities and at the RSM. We interviewed Elders and Knowledge Keepers from Pelican Narrows and Sturgeon Lake and recorded their oral stories; collected Indigenous artifacts in these communities and selected samples from RSM collections for physical measurements at the Scanning Electron Microscope Laboratory of the University of Alberta, Saskatchewan Isotope Laboratory of the University of Saskatchewan and André E. Lalonde Accelerator Mass Spectrometry Laboratory of the University of Ottawa. Then we carried out the statistical analysis of the obtained data. The preliminary results of the project were presented to community members.

Human choroid plexus in a patient with intracranial hypertension and hydrocephalus: scanning electron microscopy

Introduction The daily production of 500 ml of cerebral spinal fluid (CSF) is largely carried out by the choroid plexus (CP), which is located in the cerebral ventricles. In recent years, much importance has been given to studies of the choroid plexus for a better understanding of the mechanism of headache associated with idiopathic intracranial hypertension or other etiologies. A few drugs acting on the CP to reduce CSF secretion are used to treat headache in patients with benign intracranial hypertension. Objectives To show scanning electron microscopy images of CP obtained from a pediatric patient with hydrocephalus. Method The CP sample was collected during a surgical procedure of a third ventriculostomy and CP coagulation as a treatment for hydrocephalus in a 2-year-old boy. A sample of uncoagulated CP (5 mm) was removed and immediately immersed in glutaraldehyde solution and refrigerated at 4°C. In the electron microscopy laboratory of UFPE, the sample. The sample was fixed with Karnovsky solution (2.5% glutaraldehyde, 4.0% formaldehyde and 0.1M sodium cacodylate buffer), later postfixed with 1% osmium tetroxide, dehydrated in an increasing series of alcohol (30%, 50%, 70%, 90% and 100%) and carried out the critical point for total drying. The samples were placed in metallic stubs containing a double-sided carbon tape and then metalized with gold-palladium in the Ion Sputter JFC-1100 (JEOL) equipment. Images were performed using the EVO LS-15 scanning microscopy (ZEISS). Results We present images of CP segments at 20,000 to 40,000x magnification, showing surface projections suggesting microvilli. Due to ethical issues, we do not have a control sample of healthy individuals for comparative purposes. Conclusion To the best of our knowledge, this is the first time human CP images of a patient with hydrocephalus are presented using scanning electron microscopy. Keywords: Choroid plexus, Headache, Intracranial hypertension, Hydrocephalus, Microscopy.

MO053: Are Renal Prognostic Markers Different in Patients with Exostosin 1/2 Positive or Negative Membranous Lupus Nephritis? A Single Center Experience in Turkey

Abstract BACKGROUND AND AIMS Recently, prognostic importance of antibodies in primary membranous nephropathy is well defined. However, knowledge about antibodies related to disease activity in membranous lupus nephritis (MLN) is limited. Exostosin1 and exostosin2 (EXT1/2) are novel antibodies detected in MLN, and further studies are needed for the prognostic significance of them as a marker. The aim of this study is to determine whether renal prognostic markers differ between EXT1/2 positive and negative MLN patients. METHOD This is a single-center, retrospective and observational study. Class 5 MLN patients aged 18 or older were included in the study. Clinical characteristics, results of biochemical analysis and histopathological examination were recorded from 6 to 18 months. The end-point of the study was determined as complete remission, partial remission, need for renal replacement therapy (RRT) and exitus. Group 1: EXT1/2 positive patients. Group 2: EXT1/2 negative patients. Light, immunofluorescent and electron microscopy findings and laboratory parameters were evaluated in EXT1/2 positive and negative patients. RESULTS Group 1: n = 11 patients. Group 2: n = 7 patients. The main demographic, biochemical and histopathological characteristics of the groups were shown in Table 1. EXT1/2 negative patients, a higher number of sclerotic glomeruli, interstitial fibrosis and a higher amount of proteinuria were present than positive patients (P < 0.05). EXT1/EXT2 negative patients had significantly lower C3 and C4 levels than positive patients (P < 0.05). EXT1/EXT2 negative patients had a higher RRT or exitus rate within 18 months than positive patients (P < 0.05). In contrast, EXT1/EXT2 positive patients had a higher rate of achieving complete or partial remission within 18 months than negative patients (P < 0.05). The effect of immunosuppressive drug use other than steroids and glomerulosclerosis remission was statistically significant (P = 0.04 and P = 0.011). CONCLUSION EXT1/2 negative class 5 LMN patients have more poor renal prognostic indicators than positive patients. RRT or exitus rate is higher in EXT1/EXT2 negative patients than positive patients. However, more randomized controlled studies are needed.

Outbreak of paramyxovirus in Chestnut-bellied Seed-Finch (Sporophila angolensis). Transmission electron microscopy diagnosis / Surto de paramixovirose em curiós Sporophila angolensis). Diagnóstico por microscopia eletrônica de transmissão

The Chestnut-bellied Seed-Finch (Sporophila angolensis) is a bird constantly sought after as a cage bird, which is the main threat and the cause of its disappearance from the most inhabited regions of the country. Paramyxovirus, present in most breeding sites and Ecological Parks, is the disease that most affects birds with a high mortality rate, causing significant losses, both economic and to nature. Avian paramyxoviruses belong to the family Paramyxoviridae, which includes 21 serotypes (APMVs-1-21). Serotypes 2 and 3 are the ones that most infect passerines. Type 2 causes respiratory disorders, while the disease caused by serotype 3 is characterized by pancreatitis, conjunctivitis, vomiting, diarrhea, dyspnea, dysphagia, pancreatitis, CNS symptoms, and death. Free-ranging birds can act as viral reservoirs and spread the disease when subjected to stress factors. During the period of 2009, during the illegal commercialization of Brazilian birds, 218 Chestnut-bellied Seed-Finch were apprehended and sent to the CRAS (Wild Animals Recovery Center) of the Tietê Ecological Park. Of these, 112 arrived dead and the rest died during the year. About 35 birds were sent to the Electron Microscopy Laboratory of the Biological institute, for research on viral agents. During the necropsy, it was observed that the intestines were dilated, containing yellowish and watery stools. Small intestine fragments were processed by negative staining (rapid preparation), antigen immunolabeling with colloidal gold particles and resin embedding. Under the transmission electron microscope, using the negative staining technique, pleomorphic, rounded or elongated, enveloped paramyxovirus particles containing helical herring-bone like nucleocapsid, measuring between 100 and 500 nm in diameter, were visualized in the samples of all birds. The antigen-antibody reaction was enhanced by colloidal gold particles. Ultrathin sections of the small intestine revealed the presence of nuclei with marginalized chromatin, containing intranuclear inclusions; amorphous intracytoplasmic inclusions, formed by helical nucleocapsids, complete particles measuring from 100 to 250 nm and incomplete ones, budding from the plasma membrane.

Detection of avipoxvirus in a cutaneous lesion of a swallow tanager (Tersina viridis) by transmission electron microscopy / Detecção de avipoxvírus em lesões cutâneas de saí-andorinha (Tersina viridis) por microscopia eletrônica de transmissão

Avianpox is a highly contagious disease that affects worldwide both commercial and wild birds. The etiologic agent is a poxvirus, belonging to the Poxviridae family and Avipoxvirus genus. The disease manifests itself in three ways, cutaneous, diphtheric and septicemic. In June 2007, during the illegal commercialization of birds, a Swallow tanager (Tersina viridis) was apprehended by the Forestry Police, among other birds, which was sent to the Tietê Ecological Park, São Paulo, SP, Brazil. After one month, the bird presented skin lesions on its legs, anorexia, emaciation, mobility difficulties, diarrhea, dehydration and death, and was sent to the Electron Microscopy Laboratory of the Biological Institute, São Paulo, SP, Brazil, to search for viral agents. Fragments of skin lesions and crusts were collected and processed for transmission electron microscopy using the techniques of negative staining (rapid preparation) and immunocytochemistry (immunollabelling with colloidal gold particles). Under the transmission electron microscope using the negative staining technique, a large number of typical poxvirus particles ovoid in shape, showing irregular arrangement of the tubules on the outer membrane, some enveloped, measuring an average of 240 nm in length x 200 nm in diameter, were seen in suspension of skin lesions or crusts. . In the immunocytochemistry technique the antigen-antibody reaction was strongly enhanced by the dense particles of colloidal gold on poxviruses.

Characterization of Macro Mechanical Properties and Microstructures of Cement-Based Composites Prepared from Fly Ash, Gypsum and Steel Slag

Using solid wastes (SWs) as backfilling material to fill underground mined-out areas (UMOAs) solved the environmental problems caused by SWs and reduced the backfilling cost. In this study, fly ash (FA), gypsum and steel slag (SS) were used to prepare cement-based composites (CBC). The uniaxial compression, computed tomography (CT) and scanning electron microscope (SEM) laboratory experiments were conducted to explore the macro and micromechanical properties of CBC. The findings showed that the uniaxial compressive strength (UCS) of CBC with a curing time of 7 d could reach 6.54 MPa. The increase of SS content reduced the UCS of CBC, while the gypsum and FA content could increase the UCS of CBC. Microscopic studies have shown that the SS particles in CBC have noticeable sedimentation, and the increase of SS content causes the failure mode of CBC from tensile to tensile-shear. These research results can provide a scientific reference for the preparation of backfilling materials.

Бактерійна адгезія на клітини епітелію піднебінних мигдаликів при інфекційному мононуклеозі

Aim. To evaluate the features of pharynx tonsils mucous membrane colonization by pathogenic and opportunistic microorganisms using electron microscopic examination of pharynx palatine tonsils epithelium in patients with infectious mononucleosis and acute streptococcal tonsillitis. Materials and Methods. Two patients, i.e. a patient P. (12 years old) with a confirmed diagnosis of infectious mononucleosis, and patient A. (8 years old) with acute streptococcal tonsillitis diagnosis were examined. A bacteriological examination of mucus and epithelium scraping from the surface of the pharyngeal tonsils was conducted. Tissue samples were examined in the electron microscopy laboratory of the Lviv National University. Results and Discussion. 36 tonsils epithelial tissue micro preparations of patient A. and 41 micro preparations of patient P. were studied. Streptococcus pyogenes, Str. pneumoniae, Str. viridans, Сandida albicans, as well as non-pathogenic bacteria, Diphtheroides sp., Neisseria sp., and Corynebacterium spp. were identified as result of the bacteriological examination of mucus from the surface of the pharyngeal tonsils of the patient P.. Staph. aureus, Str. Viridans, and Str. pneumoniae were identified during a bacteriological examination of the patient A. Eosinophils with a two-segmented nucleus, specific granularity, phagocytosed spherical bacteria in the cytoplasm were detected during the histological examination of the materials taken from the surface of the patient with acute tonsillitis tonsils. Research shows that bacteria accumulate not only in the structure of extracellular detritus. Numerous bacteria accumulations were found in the cytoplasm of the epithelial cells in the patient with infectious mononucleosis also. The cell's shape resembles a bunch of grapes. Conclusions. The electron microscopic examination showed differences in the coccal flora localization. The extracellular localization of bacteria in the patient with acute bacterial tonsillitis, and intraepithelial presence of the bacteria in the patient with tonsillitis during infectious mononucleosis were found.

The investigation of bacterial adhesion of palatine tonsils epithelial cells in patient with infectious mononucleosis

Aim. To evaluate the features of pharynx tonsils mucous membrane colonization by pathogenic and opportunistic microorganisms by using the pharynx palatine tonsils epithelium electron microscopic examination of the patients with infectious mononucleosis and acute streptococcal tonsillitis.
 Materials and methods. Two patients – patient P., 12 years old with a confirmed diagnosis of infectious mononucleosis and patient A., 8 years old with confirmed acute streptococcal tonsillitis. The patients were taken a bacteriological examination of the mucus and epithelium scraping from the surface of the pharyngeal tonsils. Tissue samples were examined in the electron microscopy laboratory of the Lviv National University.
 Results and discussion. 36 tonsils epithelial tissue micro preparations of patient A. and 41 micro preparations of patient P. were studied. Streptococcus pyogenes, Str. pneumoniae, Str. viridans, Сandida albicans, as well as non-pathogenic bacteria: Diphtheroides sp., Neisseria sp., Corynebacterium spp. were identified as result of the patient P. bacteriological examination of mucus from the surface of the pharyngeal tonsils. Staph. aureus, Str. viridans, Str. pneumoniae were identified during a bacteriological examination of patient A. Eosinophils with a two-segmented nucleus, specific granularity, phagocytosed spherical bacteria in the cytoplasm were detected during the histological examination of the materials taken from the surface of the patient’s tonsils with acute tonsillitis. Research showed that bacteria were accumulated not only in the structure of extracellular detritus. Numerous bacteria accumulations were also found in the cytoplasm of the epithelial cells in the patient with infectious mononucleosis. The cell's shape looked like a bunch of grapes.
 Conclusions. The electron microscopic examination showed differences in the coccal flora localization: the extracellular localization of bacteria in the patient with acute bacterial tonsillitis and intraepithelial presence of the bacteria in the patient with tonsillitis during infectious mononucleosis were found.

NexusLIMS: A Laboratory Information Management System for Shared-Use Electron Microscopy Facilities.

This work introduces NexusLIMS, an electron microscopy laboratory information management system designed and implemented by the Office of Data and Informatics and the Materials Science and Engineering Division at NIST for a multi-user electron microscopy co-op facility. NexusLIMS comprises network infrastructure, shared information technology resources, a custom software package to harvest and extract experimental information and construct experimental metadata records, and an intuitive web-based user-facing interface for searching, browsing, and examining research data. These metadata records conform to the Nexus Experiment schema, which is introduced in this work. The NexusLIMS suite of tools requires minimal input and adjustments to user behavior, instead relying on existing organizational procedures and the collection of information from a multitude of sources to construct a complete picture and record of a research experiment. The underlying infrastructure and design considerations for a multi-user data management system are discussed. The core codebase for NexusLIMS is made publicly available alongside this work, and its modular design encourages the adaptation of the presented methods at other research organizations.

Probelab ReImager: An Open-Source Software for Streamlining Image Processing in an Electron Microscopy Laboratory

Providing high-quality electron images and hyperspectral X-ray maps is a focus of many modern electron microscopy laboratories. Nevertheless, further image processing and annotations are often needed to prepare them for publications and reports. For multi-user facilities, accessibility to processing software can be a limitation either through license costs or availability of processing stations. Open-source software running on multiple platforms allows for post-acquisition data processing in-lab or on user-owned devices. We developed Probelab ReImager to supersede our vendor-supplied acquisition software's exportation by being efficient and highly customizable. This article describes its main features and capabilities.

Ceramic Glazed Painted Eggs Production Technology Based on Kyiv Archaeological Materials

Glazed painted eggs were one of the most vivid manifestations of Ancient Rus’ ceramic art. Nevertheless, today there are still unsolved problems in study of this category of archaeological material. It concerns technological aspects. These articles were made of ordinary pottery loam in a band way. They were formed hollow and contained a small ceramic ball in the inner part. After the molding they were fired, then covered with transparent glaze, which served as a background. In most cases green and fulvous glaze was used. Then, by using a special tubular instrument the painting was made with glaze of another colour, usually yellow and green, and the item was heated up again in the kiln in order to smooth the surface. Paintings were not multifarious. There were three main types of designs: ordinary stripes, which surrounded the item several times; unsystematic stripes that were put lengthwise and athwart; and also braces, which were the most common ornament. The question of the Ancient Rus’ glaze, which covered the glazed painted eggs, was raised in the 1960-ies by T. Makarova. However, today, on the basis of our collection and in the context of significant changes in the field of science, we have succeeded in clarifying and extending the known facts. The Nanomedtech Electron Microscopy Laboratory with the help of a Tescan Mira 3 LMU scanning microscope and the Gatan Pecs 682 precision etching and coating system was kindly performed with spectral analysis. Spectral analysis confirmed that Ancient Rus’ glaze had a lead-silica composition with a high content of tin. Brown colour was obtained by iron oxide, yellow by lead, green by copper. A general glaze formula was developed, which should consist essentially of Pb and Si oxide. On the example of two samples that have the same yellow colour, the difference in the quantitative composition of the components and their percentage can be seen that, by-turn, confirms the view of the relative dimensions of component measurement. The analysis of a small amount of material allowed describing the technology of painted eggs manufacturing.

Formation of wide-blocky calcite veins by extreme growth competition

Liassic limestones on the coast of Somerset in the UK contain dense arrays of calcite microveins with a common, but poorly understood microstructure, characterized by laterally wide crystals that form bridges across the vein. We investigated the mechanisms of formation and evolution of these ‘wide-blocky’ vein microstructures using a combination of high-resolution analytical methods, including virtual petrography, optical cathodoluminescence and scanning electron microscopy techniques (e.g. energy-dispersive X-ray spectrometry, back-scattered electron imaging, cathodoluminescence and electron back-scattered diffraction), laboratory experiments and multiphase field modelling. Our results indicate that the studied veins formed in open, fluid-filled fractures, each in a single opening and sealing episode. As shown by the optical and electron back-scattered diffraction images, the vein crystals grew epitaxially on grains of the wall rock and we hypothesize that their growth rates differed depending on whether the crystals were on a wall rock grain substrate that fractured intergranularly (slow growth rates) or transgranularly (rapid growth rates). Our multiphase field models support this hypothesis, showing that wide, blocky crystals only form where there are significant differences in the growth rate and are dependent on the type of seed grain. These results provide strong evidence for extreme growth competition, a process that we propose controls vein-filling in many micritic carbonate reservoirs, as well as demonstrate that the characteristics of the fracture wall can affect the filling processes in syntaxial veins. Supplementary material: The description and images of the studied thin sections are available at https://doi.org/10.6084/m9.figshare.c.5172371 . High-resolution optical microscopy mosaics (under plane-polarized- and crossed polarized light) of the thin section collection in PetroScan file format are available on request from the authors.

Diagnostic signs of morphological and anatomical structure of Medicago falcata L. raw materials

Topiciality. Flora of Azerbaijan has a diverse vegetation cover and is rich in choice. However, today these sources of raw materials are not fully used to meet the needs of medicine and national economy. We conducted a resource reconnaissance survey of species of the genus Medicago in Azerbaijan and identified regions of mass growth. New natural habitats are located in different environmental conditions and it is possible to change the diagnostic features of the morphological and anatomical structure of plant materials, as well as the chemical composition. The combination of additional morphological and anatomical features allows a more accurate determination of the authenticity of the raw material. Aim. To study the diagnostic signs of morphological and anatomical structure of Medicago falcata L. raw materials from the flora of Azerbaijan. Materials and methods. Plant samples were fixed in a solution made in 0.1 M phosphate buffer (pH = 7.4), containing 2.5 % glutar-aldehyde, 2 % paraformal-aldehyde, 4 % sucrose, and 0.1 % picric acid. The fixed materials have been submitted to the Electron Microscopy Laboratory of the Azerbaijan Medical University for study by electron microscopy. Samples have been postfixed in 1 %-osmium tetraoxide solution prepared in phosphate buffer (pH 7.4) within two hours after being left in the same fixer for one day. Araldite-Epon blocks made from materials using general methods adopted in electron microscopy. The semi-thin (1-2 µm) sections from the blocks taken on a EM UC7 (Leica, Germany) ultramicrotome, stained with methylene blue, azure II, and basic fuchsin were observed under microscope Primo Star (Zeiss, Germany) and images of required parts were shot with EOS D650 (Canon, Chine) digital camera. Results and discussion. Multicellular trichome covered on both leaf sides. Adaxial epidermis a sheet plate have sinuous cells, and abaxial epidermis have differed slightly sinuous cells with clear-shaped walls. In leaflet anatomy of plant vascular bundles are of the collateral type. Stomatas are located from both sides of a leaf. Stomata belong to anisocytic structure. The epidermis of the stem consists of elongated thick-walled cells with anisocytic stomata. The calyx is densely covered by trichomes. The outer epidermis of cross-section of the keel petals cells is the wing horns cells form. And inner epidermis consists of oval cells. Mesophyll cells inside the leaf consist of loose spongy cells on the underside with several conducting bundles (dorsoventral). Conclusions . Morphological and anatomical studying of raw material Medicago falcata L. has shown, that there are prominent features of a structure:four large conductive bundles are located on the cross section of the stem at the corners of the ribs on two sides. Between angular conductive bundles, there are three conductive bundles, and the other two sides are located along one conductive bundle. Therefore, M. Falcata L. differs from M. Sativa L. The epidermis of the stem consists of elongated thickened-walls cells with anisocytic stomata. The indicated diagnostic morphological and anatomical characters could be used in the compilation of a monograph and in identifying plant materials on the raw material of Medicago falcata L.

Syndecan-1 (CD138) immunohistochemical expression patterns in lupus nephritis; reflections on different clinicopathological parameters

Introduction: Syndecan 1 (SCD1) is a lectin expressed at the surface of renal tubular epithelial cells and plasma cells. In epithelial cells, cell surface syndecan1 is cleaved by inflammation-induced proteases (eg, ADAMTS, MMP), since loss of cell surface syndecan1 is associated with higher susceptibility to cell damage. Objectives: To explore a potential additional value of SCD1 immunohistochemical expression in lupus nephritis specimens of different ISN/RPS classes and NIH activity and chronicity indices. Patients and Methods: This retrospective study included 50 renal biopsy specimens diagnosed as lupus nephritis at the pathology laboratory, and electron microscopy (EM) laboratory of Ain-Shams University specialized hospitals. Data were collected from records as personal data, medical history and laboratory results including serum creatinine and proteinuria. Immunohistochemical expression of syndecan-1 was evaluated in renal tubular epithelial cells (TECs) followed by correlation with different clinicopathological parameters. Results: Fifty renal biopsy specimens with lupus nephritis including 14 cases of class II, 4 cases of class III, 20 cases of class IV and 12 cases of class V were re-evaluated. The mean serum creatinine was 1.57 ± 0.67 mg/dL. Nine cases (18%) were negative for proteinuria, while 41 cases (82%) were presented with proteinuria with a mean of 1.5 ± 0.9 g/24 h. There was no statistically significant difference in the percentage of SCD-1 expression with different lupus classes. Serum creatinine and albumin showed a statistically significantly different correlation with semiquantitative score of SCD-1 expression. The highest value of creatinine detected with score 1 of SCD-1 expression (P=0.038) and the highest value of urinary albumin was recorded with score 1 of SCD-1 expression. Accordingly, the lowest mean of urinary albumin recorded in SCD-1 score 3 (P<0.001). There was a weekly negative association between loss of SCD-1 expression and increased NIH activity and chronicity indices. Conclusion: Loss of syndecan immunohistochemical expression in renal TECs in lupus nephritis is highly associated with proteinuria and elevated serum creatinine and can be used as a predictive marker for disease severity and progression.

Volume-CLEM: a method for correlative light and electron microscopy in three dimensions.

Generation of three-dimensional (3D) data sets from serial sections of tissues imaged by light microscopy (LM) allows identification of rare structures by morphology or fluorescent labeling. Here, we demonstrate a workflow for correlative LM and electron microscopy (EM) from 3D LM to 3D EM, using the same sectioned material for both methods consecutively. The new approach is easy to reproduce in routine EM laboratories and applicable to a wide range of organs and research questions.

Evaluation of a TEM based Approach for Size Measurement of Particulate (Nano)materials.

An approach for the size measurement of particulate (nano)materials by transmission electron microscopy was evaluated. The approach combines standard operating procedures for specimen preparation, imaging, and image analysis, and it was evaluated on a series of certified reference materials and representative test materials with varying physical properties, including particle size, shape, and agglomeration state. The measurement of the median value of the minimal external particle diameter distribution was intra-laboratory validated. The validation study included an assessment of the limit of detection, working range, selectivity, precision, trueness, robustness, and ruggedness. An uncertainty that was associated to intermediate precision in the range of 1–7% and an expanded measurement uncertainty in the range of 7–20% were obtained, depending on the material and image analysis mode. No bias was observed when assessing the trueness of the approach on the certified reference materials ERM-FD100 and ERM-FD304. The image analysis method was validated in an inter-laboratory study by 19 laboratories, which resulted in a within-laboratory precision in the range of 2–8% and a between-laboratory precision of between 2% and 14%. The automation and standardization of the proposed approach significantly improves labour and cost efficiency for the accurate and precise size measurement of the particulate materials. The approach is shown to be implementable in many other electron microscopy laboratories.

The Necessity of Standards in an ISO 17025 Accredited Bioforensic Electron Microscopy Laboratory

The Necessity of Standards in an ISO 17025 Accredited Bioforensic Electron Microscopy Laboratory