Journal of Histotechnology最新文献

Lupus nephritis (LN) is a severe manifestation of systemic lupus erythematosus (SLE), characterized by marked histological heterogeneity and high interobserver variability in the evaluation of renal biopsies. Digital morphometry has emerged as an objective and reproducible tool that enables precise quantification of tissuelesions, thereby enhancing diagnostic accuracy and prognostic assessment. To explore its application in LN, a systematic review was conducted following PRISMA guidelines, including observational studies in adults, children, and animal models with biopsy-proven LN in which digital morphometry was applied. Searches were performed inPubMed, Embase, Scopus, Web of Science, Cochrane, and Google Scholar. From 376 identified records, 46 studies fulfilled inclusion criteria. Among these, 30 analyzed the glomerularcompartment, 22 the tubulointerstitial, and the vascular. Both manual techniques and image analysis software such as ImageJ, ImagePro-Plus, and QuPath were used. In the glomerular compartment, parameters including cellularity, mesangial expansion, proliferation, and immune deposits were assessed. Tubulointerstitial studies measured fibrosis, tubular atrophy, and immune Infiltrates, while vascular analyses examined intimal thickening and complementactivation. Across compartments, morphometry consistently outperformed conventional visual evaluation, particularly in fibrosis quantification and prediction of renal prognosis. Overall, digital morphometry represents a valuable method for the comprehensive assessment of LN renal biopsies. Its application provides greater precision in characterizing renal damage and offers advantages interms of objectivity, reproducibility, and prognostic value. Standardization of morphometric approaches, together with integration into artificial intelligence-based tools, could optimize diagnostic accuracy and therapeutic strategies in LN.

Annotated pathology datasets are the cornerstone for developing computational pathology. However, the intrinsic complexity of pathology data often results in a scarcity of large, manually annotated datasets. To address this challenge, a rapid, accurate, and automatic method for annotating tumor cells is essential for advancing the field of computational pathology. Here, we introduce a novel annotation technology. In our approach, Hematoxylin-eosin (H&E) slides were first digitized and preserved. These H&E slides were then faded and re-stained using multiple biological technologies (MBT) to identify specific biomarkers. The re-stained MBT slides were subsequently scanned again to create new digital images. The original and re-stained digital images underwent a registration and segmentation process to ensure that all minute structures in both images aligned perfectly. The staining results from the MBT slides, referred to as2 label maps, were extracted and transferred back onto the H&E slides, and the data were merged. This method allows for precise and efficient annotation of all tumor cells, including those expressing specific biomarkers, as well as the components of the tumor microenvironment. By rapidly generating high-quality, high-volume datasets, this innovative method significantly enhances the ability of AI approaches to analyze and interpret pathology data. Consequently, it supports the development of highly accurate diagnostic, prognostic, and predictive decision-making systems in the field of computational pathology.

Toxicologic pathology is undergoing a digital transformation, with advances in imaging and computational methods enabling automation of traditionally manual workflows. Central to these digital workflows is the generation of high-quality whole slide images (WSIs), where one key determinant of image quality is focus sharpness. To address this, we have integrated a pair of productionalized computational models - 'MiQC' (Microscopic Quality Control) - into our routine image QC workflows. MiQC combines Local Binary Patterns (LBP) and DeepFocus-based deep learning algorithms to detect and quantify out-of-focus regions in WSIs. Subsequent to scanner-based focus metric assessment, MiQC further screens WSIs and supports technician review by generating heatmaps that highlight problematic areas. Even WSIs with scanner focus scores of 98-99% can contain unacceptable blur, which MiQC helps identify. Using this system, 85-95% of WSIs are approved without further intervention, and technician review time is reduced by nearly 50%. Compared to fully manual review, MiQC has doubled our throughput of QC'd slides per hour. This efficiency gain has accelerated the expansion of our high-quality WSI repository and provides a scalable, reproducible framework for enhancing image QC in toxicologic and broader digital pathology applications. MiQC supports higher throughput and integration of automated image analysis pipelines, laying the groundwork for robust downstream computational pathology workflows.

Decalcification is an important step in histology laboratories to allow mineralized tissue samples to be trimmed and sectioned easily. Many decalcifying solutions have a rapid onset of action in softening tissues but alter protein structure and morphology, while others preserve protein integrity but are less efficient. The ideal decalcification protocol allows for rapid and cost-effective processing and precise evaluation of microscopy and antigen-based immunostaining such as immunohistochemistry. In our study, mouse tissues were decalcified with three commercially available solutions to identify the product that best meets those criteria. Immunocal^TM (StatLab), Epredia^TM, and Rapid-Cal^TM (StatLab) decalcification agents were tested on formalin-fixed, paraffin-embedded CD-1 mouse femur, skull, and sternum samples. Multiple metrics including ammonium oxalate turbidity and radiography were used to assess stages of bone demineralization. Tissues were routinely processed, embedded in paraffin, and sectioned at 5 µm for H&E staining and CD3, CD31, and Iba1 immunostaining. Each tested sample represented a decalcification product and time (4, 6, 12, 24, 30, 48, 73 h). Samples were assessed by radiolucency on X-ray and gross bone pliability prior to histologic processing, followed by histopathologic scoring for completeness of demineralization, preservation of tissue architecture, and antigenicity of tissue. All three commercially available decalcifying solutions are sufficient for rapid decalcification with preservation of tissue integrity, cellular detail, and immunogenicity.

Inorganic mercury (Hg) and methylmercury (MeHg) have emerged as global pollutants owing to their long-term environmental stability and bioaccumulation. These heavy metals enter aquatic systems via industrial emissions, coal combustion, and natural processes, posing a serious threat to ecosystems and human health. This study assesses the impact of Hg²⁺ and MeHg on the growth and development of brine shrimp (Artemia franciscana) nauplii by analyzing the histopathological effects on their tissues. Brine shrimp nauplii from the Bohai Bay in China were selected as the study subjects and exposed to 1 μmol/L solutions of HgCl₂ and MeHgCl. Tissue sections were continuously taken at different immersion times. After staining with the standard hematoxylin-eosin (HE) method, the tissue morphology of brine shrimp nauplii under different forms of Hg stress was observed under a light microscope. The results showed that MeHg exhibited significantly greater toxicity to brine shrimp nauplii than Hg²⁺. Under the same exposure time, the MeHg group exhibited more pronounced epithelial cell damage, nuclear material disorder, and nucleoplasm diffusion outside the nucleus than the HgCl₂ group. This finding provides an important theoretical support for further research into the toxicological mechanisms of MeHg and Hg²⁺, and highlights that the toxic effects of methylmercury on aquatic organisms.

Amyloidosis encompasses a spectrum of rare disorders characterized by extracellular amyloid deposition. Achieving an accurate early diagnosis of systemic amyloidosis necessitates biopsy-specific pathological evaluation. Formalin-fixed, paraffin-embedded liver biopsy specimens were examined using Congo red staining, electron microscopy, immunohistochemistry (IHC), immunofluorescence, and Congo red-assisted laser microdissection with mass spectrometry (LMD/MS). Failure Mode and Effects Analysis (FMEA) was employed for risk mitigation and quality control. Classical Congo red staining exhibited brick-red coloration, enhanced alkalinization, reduced permanganate staining, and characteristic apple-green birefringence under polarized light. Trypsin-digested IHC demonstrated kappa light chain positivity and lambda negativity, with improved background clarity compared to other retrieval methods-results concordant with electron microscopic colloidal gold staining, albeit with higher tissue consumption. Congo red-polarized microscopy permitted direct amyloid deposit localization. Subsequent LMD/MS identified immunoglobulin kappa light chain as the pathogenic precursor protein, though at increased expense. Congo red staining under polarized light remains the cornerstone technique for amyloid detection. LMD/MS provides superior specificity in amyloid typing relative to IHC, immunofluorescence, or electron microscopy, proving particularly advantageous for limited samples. Traditional methods remain valuable for validation when tissue is abundant. Histopathological assessment continues to be the diagnostic gold standard for hepatic amyloidosis; systematic integration and analytical refinement of these techniques are imperative for enhancing diagnostic accuracy.

Placentas are temporary organs needed to support a developing embryo and arise from both embryonic and maternal tissues. Calcifications of tissues outside of bone and teeth mineralization are often a sign of tissue damage and impaired organ function. Placental calcifications have been described previously in the literature and usually increase in normal pregnancies as the placenta ages, but they have also been associated with the potential for fetal distress. This study utilized 139 placental tissues from singleton control placentas (51), fused twin placentas (48), and non-fused twin placentas (40) with weights over 320 grams (third trimester) and similar maternal ages to determine which placental type(s) exhibited the largest density of calcifications using the von Kossa stain for calcium salts. The study found there were no differences in calcification densities among placenta types, suggesting that twin pregnancies do not experience additional placental stress risk from calcifications. Importantly, the finding that nearly all third-trimester placentas contained calcifications when systematically evaluated indicates that calcification may represent a normal maturational process rather than a pathologic sign of distress. These results help to caution the interpretation of placental calcifications and may provide reassurance to patients and providers managing twin pregnancies.

Gomori methenamine silver (GMS) stains are commonly utilized to exclude fungal esophagitis in esophageal biopsies. Our laboratory protocol for special stains, including GMS, stipulates that two levels be performed for each stain. We retrospectively reviewed 127 esophageal biopsies which had GMS stains performed; there were 62 GMS-positive and 65 GMS-negative cases. Sixty-one of the positive cases (98%) had fungal organisms present on all levels. Only one case showed fungal elements on the first, but not the second, level. Given national laboratory staff shortages, performing levels for special stains may be unnecessary, inefficient, and time consuming in the surgical pathology laboratory.

Glial fibrillary acidic protein (GFAP) immunohistochemistry staining is specific to glial cell intermediate filaments. GFAP has been reported in some diseases related to injury of central nervous system. The actual normal biological function of GFAP in the retinas of some vertebrates is still under investigation. The current study examines the distribution of GFAP in the retinas of various species of amphibian, reptilian, birds, and mammals by immunohistochemical technique. The results indicated that GFAP localization in the retina is related to specific species. Unique, homogeneous GFAP labelling was observed in Chalcides ocellatus retina, which was different from other studied reptiles (Trachemys scripta elegans, Uromastyx aegyptius, Ptyodactylus hasselquistii, Acanthodactylus boskianus, Scincus mitranus). Low GFAP labelling was noted in the retinas of birds and some species of the studied mammals (Jaculus jaculus, Rousettus egyptiacus, and Mesocricetus auratus). However, other studied mammals (Mus musculus, Acomys russatus, Paraechinus aethiopicus) showed different intensities of GFAP expression. The study finds that GFAP helps to clarify the potential function of Müller cells in the regeneration process in different species.