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Background: Formalin-fixed, paraffin-embedded (FFPE) tissues are a valuable resource for clinical and basic science research. Paraffin blocks and the resulting unstained sections (USS) are often st...

Conventional clinical histopathology analysis is based on bright-field microscopy of thinly sliced tissue specimens. Although bright-field microscopy is a simple and robust method for examining microscope slides, preparing the slides needed is a lengthy and labor-intensive process. Slide-free histopathology technologies address this problem by directly imaging intact, minimally processed tissue samples using advanced optical imaging systems, bypassing the extended workflow now required for preparation of tissue sections. This manuscript explains the technical basis of slide-free microscopy, reviews common slide-free optical microscopy techniques, and discusses the opportunities and challenges involved in clinical implementation.

Observing chromosomes is a time-consuming and labor-intensive process, and chromosomes have been analyzed manually for many years. In the last decade, automated acquisition systems for microscopic images have advanced dramatically due to advances in their controlling computer systems, and nowadays, it is possible to automatically acquire sets of tiling-images consisting of large number, more than 1000, of images from large areas of specimens. However, there has been no simple and inexpensive system to efficiently select images containing mitotic cells among these images. In this paper, a classification system of chromosomal images by deep learning artificial intelligence (AI) that can be easily handled by non-data scientists was applied. With this system, models suitable for our own samples could be easily built on a Macintosh computer with Create ML. As examples, models constructed by learning using chromosome images derived from various plant species were able to classify images containing mitotic cells among samples from plant species not used for learning in addition to samples from the species used. The system also worked for cells in tissue sections and tetrads. Since this system is inexpensive and can be easily trained via deep learning using scientists’ own samples, it can be used not only for chromosomal image analysis but also for analysis of other biology-related images.

Pu‐erh tea is very popular in Southwestern China and South Asian countries and is now becoming increasingly popular in Europe due to its well‐documented beneficial effects on human health. Pu‐erh tea aqueous extracts can maintain intestinal homeostasis. However, the mechanism of its beneficial effects on intestinal flora disorder is not clear. In this study, we focused on the effects of ripe Pu‐erh tea aqueous extracts on the intestinal microbiota in an intestinal flora disorder mouse model. Physiological indexes and the tissue section staining results showed that feeding Pu‐erh tea extract could help mice regain weight and alleviate intestinal inflammation. Further assessment of the intestinal microflora found that Pu‐erh tea extract could promote the growth of intestinal probiotics and inhibit pathogenic bacteria, thereby achieving a treatment effect for enteritis. This study provides new evidence for the therapeutic effect of Pu‐erh tea., Pu‐erh tea is very popular in Southwestern China and South Asian countries, and now is becoming more and more popular in Europe because of its well‐documented beneficial effects on human health. This study explored the mechanism of Pu‐erh tea extract in promoting human health and improving intestinal flora disorder and found that Pu‐erh tea extract can alleviate the intestinal inflammation caused by antibiotics, reduce the degree of intestinal lesions, promote the growth of intestinal probiotics and inhibit pathogenic bacteria.

Li Cui,1 Zirong Zhu,2 Yiguo Feng,3 Yanfei Zhang3 1Department of Dermatology, Shaanxi Provincial People’s Hospital, Xian, People’s Republic of China; 2Department of Dermatology, Xi’an People’s Hospital, Xian, People’s Republic of China; 3Department of Dermatology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xian, People’s Republic of ChinaCorrespondence: Yanfei ZhangDepartment of Dermatology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xian, People’s Republic of ChinaEmail zhangyanfei723@163.comPurpose: Keratin 17 (K17) is an embryonic keratin and overexpression is seen in psoriasis, which is a hyperproliferation skin disease. Nonetheless, whether it is also highly expressed in other proliferative skin diseases remains unclear. The aim of this study is to explore the expression of K17 in cutaneous lichen planus (CLP), lichen simplex chronicus (LSC), and prurigo nodularis (PN).Methods: A total of 20 skin samples from CLP lesions, 20 from LSC lesions, 20 from PN lesions, and 10 healthy adult skin tissues were obtained. Then, the expression of K17 was analyzed using immunohistochemistry on paraffin-embedded tissue sections. Furthermore, quantitative and semi-quantitative immunohistochemical scores of K17 were independently evaluated under a microscope by 2 dermatologists.Results: Immunohistochemical analysis revealed that in normal skin, K17 was minimally expressed. Nevertheless, it was highly expressed in all epidermal layers in CLP lesions (P-value < 0.01), and negatively expressed in LSC and PN lesions (all P-value > 0.05). The average gray value of K17 in CLP was 151.153± 13.985 (P-value < 0.001), while the average values of K17 in LSC and PN were 178.720± 12.001 and 181.316± 8.920, respectively (all P-value > 0.05).Conclusion: K17 is potentially expressed in certain inflammatory skin diseases, including psoriasis and lichen planus. Besides, it is not always a marker of hyperproliferation of keratinocytes in skin diseases.Keywords: keratin 17, proliferation of keratinocytes, inflammatory skin diseases

Tissue microenvironment properties like blood flow, extracellular matrix, or proximity to immune-infiltrate are important regulators of cell biology. However, methods to study regional protein expression in the native tissue environment are limited. To address this need, we developed a novel approach to visualize, purify, and measure proteins in situ using automated spatially targeted optical microproteomics (AutoSTOMP). Here, we report custom codes to specify regions of heterogeneity in a tissue section and UV-biotinylate proteins within those regions. We have developed liquid chromatography-mass spectrometry (LC-MS)/MS-compatible biochemistry to purify those proteins and label-free quantification methodology to determine protein enrichment in target cell types or structures relative to nontarget regions in the same sample. These tools were applied to (a) identify inflammatory proteins expressed by CD68+ macrophages in rat cardiac infarcts and (b) characterize inflammatory proteins enriched in IgG4+ lesions in human esophageal tissues. These data indicate that AutoSTOMP is a flexible approach to determine regional protein expression in situ on a range of primary tissues and clinical biopsies where current tools and sample availability are limited.

As herbaria move to digitize their collections, the question remains of how to efficiently digitize collections other than standard herbarium sheets, such as wood slide collections. Beginning in September 2018, the Harvard University Herbaria began a project to image and digitize the wood slides contained in the Bailey-Wetmore Wood Collection. The primary goal of this project was to produce images of the wood tissue that could be used for specimen-level research and to make them available on the internet for remote scholarship. A secondary goal was to establish best practices for digitizing and imaging a microscope slide collection of tissue sections. Due to the size of the wood slide collection (approximately 30,000 slides), a medical histology scanner and virtual microscopy software were used to image these slides. This article outlines the workflow used to create these images and compares the results with digital resources currently available for wood anatomy research. Prior to this project, the very little of the Bailey-Wetmore Wood Collection was cataloged digitally and none of it was imaged, which made access to this unique collection difficult. By imaging and digitizing 6,605 slides in the collection, this project has demonstrated how other institutions can make similar slide collections available to the broader scientific community.

Objective: To determine the frequency of change in diagnosis and level of errors in cases referred to Armed Forces Institute of Pathology for second opinion and their impact on modifications of treatment and prognosis. Study Design: Cross sectional study. Place and Duration of Study: Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, form Mar to Oct 2017 Methodology: All the cases referred for review diagnosis were tested by applying panel of immunohistochemical markers and special stains on formalin fixed paraffin embedded tissue sections as decided on morphology. Level of errors were defined as level I error: minor discrepancy with no impact on management, level II: minor discrepancy with impact on management, level III: main category remains same but there was change/confirmation of specific diagnostic entity with an impact on management and level IV: gross changes in diagnosis with significant impact on management. Level IV was further subdivided into IV a: benign misdiagnosed as malignant, IV b: malignant misdiagnosed as benign and IV c: changes in tumor subtype. Results: A total of 100 cases, where review diagnosis was changed were included. Minor discrepancies (level I and level II) were observed in only 7% cases with little or no impact on the management. Most frequent discrepancy observed as Level III in 75% cases. Major discrepancy (Level IV errors) was noted in 18% cases. Conclusion: There were high discrepancy rates between previous diagnosis and review diagnosis. The higher use of extended panels of immunohistochemistry markers were the most likely explanations.

Characterization of tissue architecture is important to understand cell function and mechanisms in vivo. Spatial transcriptomics (ST), by recovering the transcriptome of a cell or tissue section while maintaining the two-dimensional location of the cell or tissue, can provide detailed insights into the cellular function in association with tissue architecture. Still, however, algorithmic development for ST has focused on identifying cells or spots with the same cell types located nearby and, therefore, cannot detect a tissue structure often composed of multiple cell types. Here, we present Vesalius to decipher tissue anatomy from ST data by converting transcriptomic information into a color code for image segmentation. Vesalius successfully detected tissue architecture in mouse embryo and brain from high resolution ST data by incorporating image processing algorithms. In contrast, previous spatial clustering approaches developed for low resolution ST data were unable to recover these structures. Intriguingly, Vesalius identifies genes linked to the morphology of tissue structures. In short, Vesalius is a tool to perform high-resolution in silico anatomization and molecular characterization from ST data.

Anti-fungal blue light (ABL) therapies have been studied and applied in treating various diseases caused by fungal infection. The existing work has been mainly devoted to study the effect of various light dosages on the fungal viability and on the induced cytotoxic reactive oxygen species (ROS) in the pathogens. While in vivo experimental studies have also been reported, there is still no work targeted on quantifying the effect of light on prohibiting the pathogens from invading into the deeper sites in the skin of their host. This can be attributed to the lack of methods to analyze the tissue section images, which are the main means of examining infected tissues. This work has been devoted to solve such problems, so as to improve dosimetric analyses of ABL therapies on treating fungal infections. Specifically, the invasion depth of the fungi and their ratios to the tissue in four bins at different depths inside the skin were extracted from the tissue section images. The significance of the treatment with different dosages on inhibiting the fungi was also tested by each of these depth-related metrics. The ABL experiments using 415-nm-wavelength LED light were performed on BALB/c mice, whose skin was infected by Candida albicans (C. albicans). The proposed methods were applied to the tissue sections of the experimental animals. The results clearly verified that the fluence up to 180J/cm2 can significantly prohibit the fungal infection into the skin in terms of almost all the newly proposed metrics.

Pathological detection using immunohistochemistry (IHC) has become an indispensable process in the diagnosis confirmation of various cancers. However, the production of monoclonal antibodies is always very complex, expensive and time-consuming, and the batch differences are significant due to the corporeity and health statuses of animals may be different. In this work, an aptamer-based histochemistry (aptahistochemistry) assay was developed using a DNA aptamer for specific diagnosis of clinical breast cancer tissue sections. This aptahistochemistry assay can specifically distinguish Luminal A breast cancer molecular subtype from Luminal B (HER2+), HER2-enriched, and triple-negative breast cancer molecular subtypes, as well as para-carcinoma tissue, mastitis tissue and normal breast tissue. The accuracy of this aptahistochemistry assay for the diagnosis of Luminal A breast cancer was as high as 80%, which showed a great potential for clinical pathological diagnosis applications.

RecurIndex, a multigene profiling assay, can predict the risk of local recurrence and distant metastasis in female breast cancer (FBC), but its role in male breast cancer (MBC) remains unclear. In this study, the clinicopathological data of 43 consecutive MBC patients undergoing surgeries between 2009 and 2018 were retrospectively analysed. Their paraffin-embedded tissue sections were examined by RecurIndex test which comprised 2 models: recurrence index for local recurrence (RI-LR) and recurrence index for distant recurrence (RI-DR). Of 43 patients, there were 26 low-risk and 17 high-risk patients assessed by RI-LR, while 17 low-risk and 26 high-risk patients by RI-DR. For RI-LR, tumor N stage showed statistically significant (P P = 0.033), T stage (P = 0.043) and N stage (P = 0.003). In terms of clinical outcomes, the overall survival (OS) of low- and high-risk patients stratified by RI-LR showed no statistically significant differences (P = 0.460), while high-risk patients identified by RI-DR had a significantly worse distant recurrence-free survival (DRFS) (P = 0.035), progression-free survival (PFS) (P = 0.019) and OS (P = 0.044) than low-risk patients. Overall, RI-DR can effectively predict the DRFS, PFS and OS of MBC patients and identify those at low risk of recurrence, which may serve as a potential prognostic tool for MBC.

Background. The morphopathogenesis of orofacial cleft development is only partly understood; therefore, it is important to identify factors, which possibly could be involved in it. The aim of the study was to evaluate the distribution of TGF-β1 and EGFR-containing cells in cleft affected lip mucosa.Materials and Methods. The study group included lip mucosa tissue samples from 14 patients with orofacial cleft. The control group contained 11 healthy oral mucosa tissue samples. The tissue sections were stained by immunohistochemistry for TGF-β1 and EGFR. The expression of positive structures was graded semiquantitatively. IBM SPSS 26.0 was used for statistical analysis, Spearman`s rank correlation and Mann-Whitney U tests were performed.Results. Mostly few to moderate number (+/++) of TGF-β1-containing cells was found in epithelium, also the same number of fibroblasts and macrophages was seen in the lamina propria of cleft affected lip mucosa. Meanwhile, healthy oral mucosa on average demonstrated a moderate number (++) of TGF-β1-containing epithelial cells, fibroblasts, and macrophages. A variable, mostly indistinct number of EGFR-containing cells was seen in the epithelium of cleft affected lip mucosa, meanwhile, mostly no (0) EGFR positive cells were found in the epithelium of healthy mucosa. Statistically significantly less TGF-β1-containing cells were found in the epithelium of cleft affected lip mucosa than in the healthy mucosa (U=33.000; p=0.015). Also, the lamina propria of cleft affected lip mucosa showed statistically significantly less TGF-β1 immunoreactive fibroblasts and macrophages than the healthy mucosa (U=28.500; p=0.006).Conclusions. The decreased number of TGF-β1-containing epithelial cells, fibroblasts and macrophages in cleft affected lip mucosa proves the role of problematic tissue remodelation in the cleft pathogenesis. The distribution of EGFR in cleft affected and healthy mucosa is similar and possibly does not play a role in the cleft development of humans.

The classical diagnostic principles for applying histopathology for the diagnosis of invasive fungal diseases are reviewed. Although several new molecular based techniques have recently been developed, the histopathological identification of fungal elements together with a typical tissue reaction remains the golden standard for stating a diagnosis of invasive mycosis. Therefore, and due to the risk of false negative and false positive results obtained from cultivation as well as the non-culture based diagnostic test for invasive fungal infections, an examination should always complement histopathology in the diagnosis of invasive fungal diseases. The application of molecular in situ identification techniques, i.e., immunohistochemistry and in situ hybridization, for morphologically observed fungal elements in tissue sections, has indeed improved the diagnostic accuracy of histopathology for the diagnosis of invasive fungal diseases. Because the specific molecular techniques applied in the histopathological diagnosis of invasive mycoses are directed toward specific targets, the panel of specific immunoglobulins/probes to be used on tissue sections should be directed from the histomorphology of the fungal elements as detected by conventional histopathological methods.

Single-cell RNA-seq is a tool that generates a high resolution of transcriptional data that can be used to understand regulatory networks in biological systems. In plants, several methods have been established for transcriptional analysis in tissue sections, cell types, and/or single cells. These methods typically require cell sorting, transgenic plants, protoplasting, or other damaging or laborious processes. Additionally, the majority of these technologies lose most or all spatial resolution during implementation. Those that offer a high spatial resolution for RNA lack breadth in the number of transcripts characterized. Here, we briefly review the evolution of spatial transcriptomics methods and we highlight recent advances and current challenges in sequencing, imaging, and computational aspects toward achieving 3D spatial transcriptomics of plant tissues with a resolution approaching single cells. We also provide a perspective on the potential opportunities to advance this novel methodology in plants.

Until recent years, histopathologists have analyzed tissue sections and diagnosed diseases including cancer primarily by using a microscope. The introduction of high resolution and high throughput digital scanners has enabled digitizing entire glass slides to generate high-resolution whole-slide images (WSI), de facto giving rise to the field of Digital Pathology. Since then, an increasing number of pathology laboratories have transitioned to a digital pipeline, which offers advantages such as remote diagnosis (e.g., for a second opinion), reduction of some routine work in the lab and partly reduction of physical storage. However, perhaps the most revolutionizing aspect of digital pathology is that it enables image analysis in pathology using machine learning. This field has come to be known as Computational Pathology.

Early detection of cervical lesions, accurate diagnosis of cervical lesions, and timely and effective therapy can effectively avoid the occurrence of cervical cancer or improve the survival rate of patients. In this paper, the spectra of tissue sections of cervical inflammation (n = 60), CIN (cervical intraepithelial neoplasia) I (n = 30), CIN II (n = 30), CIN III (n = 30), cervical squamous cell carcinoma (n = 30), and cervical adenocarcinoma (n = 30) were collected by a confocal Raman micro-spectrometer (LabRAM HR Evolution, Horiba France SAS, Villeneuve d’Ascq, France). The Raman spectra of six kinds of cervical tissues were analyzed, the dominant Raman peaks of different kinds of tissues were summarized, and the differences in chemical composition between the six tissue samples were compared. An independent sample t test (p ≤ 0.05) was used to analyze the difference of average relative intensity of Raman spectra of six types of cervical tissues. The difference of relative intensity of Raman spectra of six kinds of tissues can reflect the difference of biochemical components in six kinds of tissues and the characteristic of biochemical components in different kinds of tissues. The classification models of cervical inflammation, CIN I, CIN II, CIN III, cervical squamous cell carcinoma, and cervical adenocarcinoma were established by using a support vector machine (SVM) algorithm. Six types of cervical tissues were classified and identified with an overall diagnostic accuracy of 85.7%. This study laid a foundation for the application of Raman spectroscopy in the clinical diagnosis of cervical precancerous lesions and cervical cancer.

One of the main challenges in cancer surgery is to ensure the complete excision of the tumour while sparing as much healthy tissue as possible. Histopathology, the gold-standard technique used to assess the surgical margins on the excised tissue, is often impractical for intra-operative use because of the time-consuming tissue cryo-sectioning and staining, and availability of histopathologists to assess stained tissue sections. Raman micro-spectroscopy is a powerful technique that can detect microscopic residual tumours on ex vivo tissue samples with accuracy, based entirely on intrinsic chemical differences. However, raster-scanning Raman micro-spectroscopy is a slow imaging technique that typically requires long data acquisition times wich are impractical for intra-operative use. Selective-sampling Raman imaging overcomes these limitations by using information regarding the spatial properties of the tissue to reduce the number of Raman spectra. This paper reviews the latest advances in selective-sampling Raman techniques and applications, mainly based on multimodal optical imaging. We also highlight the latest results of clinical integration of a prototype device for non-melanoma skin cancer. These promising results indicate the potential impact of Raman spectroscopy for providing fast and objective assessment of surgical margins, helping surgeons ensure the complete removal of tumour cells while sparing as much healthy tissue as possible.

Optical microscopes are typically designed as versatile tools that are capable of imaging at various modalities with different resolutions in respective visual fields. A large extend of versatility causes increased complexity, size, and the cost. Imaging of tissue sections is a routine procedure in clinics and research laboratories. Tissue staining consumables, required time for staining along with the cost of the imaging systems are the limiting factors for widespread use. A multimode imaging system with enhanced resolution performance and automation is generally superfluous for the coarse resolution imaging of tissue sections. A dedicated system for label free tissue section imaging is presented. Label free imaging with high contrast is provided in form of quantitative phase. Earlier demonstrations of quantitative phase imaging of tissue sections utilized modified microscopes with different modalities for correlated imaging and motorized stages for field enlargement. Here, the system follows an off axis digital holographic imaging configuration for the acquisition of quantitative phase at single shot. The system compromises from the resolution and magnification of a traditional microscope for the size of visual field, ease of use, and relative cost due to the exclusion of sample stages. A software level stitching further enlarges the effective field of view.

Bovine mastitis, caused by Prototheca bovis, has received much attention worldwide. To investigate the status of P. bovis infection in dairy farms of Hubei, we collected 1,158 milk samples and 90 environmental samples from 14 dairy farms of Hubei, China. The isolates were identified with traditional biological methods and molecular biological techniques, and their pathogenicity was tested through mice infection experiments. Isolates from 57 milk and 20 environmental samples were identified as P. bovis. The mice infection tests proved that the isolated P. bovis could cause mastitis in mice, manifesting as severe red swelling of the mammary glands. Histopathological analysis of tissue sections showed necrosis and nodules lesions formed in the infected mice mammary tissue, accompanied by macrophage and neutrophil infiltration. These results suggested the existence of pathogenic P. bovis in dairy farms of the Hubei province, China, with brewer's grains and fresh feces possibly playing important roles in the spread of this disease.

The present study has been designed to evaluate the potent role of healing efficacy of lanolin (which is also called wool wax), is an oily product secreted by the sebaceous glands beneath the skin of sheep which is have anti-inflammatory, anti-microbial, skin protecting properties, a forty-eight mature albino rats (weighted 250-350 g) were randomly assigned to two groups; control group and eczema group which is induced by 99.9% pure acetone. By squad cotton and put it on shaved skin, after 3 days of acetone application the eczema group were divided randomly into two equal groups each group 16 rats, treat these 32 rats with glycerin and lanolin, after 24h five rats from each group were sacrificed, and remaining rats were treated with glycerin and lanolin for 72h and then sacrificed at the third day, the histopathology there is a lot of histopathological changes occur in tissue section in eczematous skin in comparison with control section; there were obvious thickness and enlargement in the length of outermost layer of skin, also there is fissures and cracks in the epidermal layer due to the dryness of skin and aggregation of inflammatory cells. And as well as keratinization of epithelial layer and changes in skin color, the results indicate there were optimum healing effect of lanolin group in compare with glycerin group after 72h, Showed back the outer most layer of the skin to its normal length, also decrease in inflammatory cells number.

The diverse composition of mammalian tissues poses challenges for understanding the cell-cell interactions required for organ homeostasis and how spatial relationships are perturbed during disease. Existing methods such as single-cell genomics, lacking a spatial context, and traditional immunofluorescence, capturing only two to six molecular features, cannot resolve these issues. Imaging technologies have been developed to address these problems, but each possesses limitations that constrain widespread use. Here we report a method that overcomes major impediments to highly multiplex tissue imaging. "Iterative bleaching extends multiplexity" (IBEX) uses an iterative staining and chemical bleaching method to enable high-resolution imaging of >65 parameters in the same tissue section without physical degradation. IBEX can be employed with various types of conventional microscopes and permits use of both commercially available and user-generated antibodies in an "open" system to allow easy adjustment of staining panels based on ongoing marker discovery efforts. We show how IBEX can also be used with amplified staining methods for imaging strongly fixed tissues with limited epitope retention and with oligonucleotide-based staining, allowing potential cross-referencing between flow cytometry, cellular indexing of transcriptomes and epitopes by sequencing, and IBEX analysis of the same tissue. To facilitate data processing, we provide an open-source platform for automated registration of iterative images. IBEX thus represents a technology that can be rapidly integrated into most current laboratory workflows to achieve high-content imaging to reveal the complex cellular landscape of diverse organs and tissues.

Anatomic pathology is an important resource for detection and exclusion of infectious diseases in tissue specimens. Detection of a microorganism (i.e. bacteria, fungi, parasite) in tissue sections is frequently the beginning of a work-up and occasionally sufficient for definitive microbiologic identification. Close correlation with cultures and ancillary testing in the microbiology laboratory is of paramount importance in arriving at a diagnosis and identify with certitude causative pathogen(s). This review will discuss the adequacy and limitations of histopathology in the diagnosis of infectious diseases, describe potential pitfalls, and discuss the appropriate use of molecular diagnostics in formalin-fixed, paraffin embedded tissues.

Current methods for spatial transcriptomics are limited by low spatial resolution. Here we introduce a method that integrates spatial gene expression data with histological image data from the same tissue section to infer higher-resolution expression maps. Using a deep generative model, our method characterizes the transcriptome of micrometer-scale anatomical features and can predict spatial gene expression from histology images alone. ispartof: NATURE BIOTECHNOLOGY vol:40 issue:4 pages:476-+ ispartof: location:United States status: published

Pathology differentiation of renal cancer types is challenging due to tissue similarities or overlapping histological features of various tumor (sub)types. As assessment is often manually conducted outcomes can be prone to human error and therefore require high-level expertise and experience. Mass spectrometry can provide detailed histo-molecular information on tissue and is becoming increasingly popular in clinical settings. Spatially resolving technologies such as mass spectrometry imaging and quantitative microproteomics profiling in combination with machine learning approaches provide promising tools for automated tumor classification of clinical tissue sections.In this proof of concept study we used MALDI-MS imaging (MSI) and rapid LC-MS/MS-based microproteomics technologies (15 min/sample) to analyze formalin-fixed paraffin embedded (FFPE) tissue sections and classify renal oncocytoma (RO, n=11), clear cell renal cell carcinoma (ccRCC, n=12) and chromophobe renal cell carcinoma (ChRCC, n=5). Both methods were able to distinguish ccRCC, RO and ChRCC in cross-validation experiments. MSI correctly classified 87% of the patients whereas the rapid LC-MS/MS-based microproteomics approach correctly classified 100% of the patients.This strategy involving MSI and rapid proteome profiling by LC-MS/MS reveals molecular features of tumor sections and enables cancer subtype classification. Mass spectrometry provides a promising complementary approach to current pathological technologies for precise digitized diagnosis of diseases.

The disconnect between preclinical and clinical results underscores the imperative for establishing good animal models, then gleaning all available data on efficacy, safety, and potential toxicities associated with a device or drug. Mini pigs are a commonly used animal model for testing orthopedic and dental devices because their skeletons are large enough to accommodate human-sized implants. The challenge comes with the analyses of their hard tissues: current methods are time-consuming, destructive, and largely limited to histological observations made from the analysis of very few tissue sections. We developed and employed cryo-based methods that preserved the microarchitecture and the cellular/molecular integrity of mini pig hard tissues, then demonstrated that the results of these histological, histochemical, immunohistochemical, and dynamic histomorphometric analyses e.g., mineral apposition rates were comparable with similar data from preclinical rodent models. Thus, the ability to assess static and dynamic bone states increases the translational value of mini pig and other large animal model studies. In sum, this method represents logical means to minimize the number of animals in a study while simultaneously maximizing the amount of information collected from each specimen.

The identification of recurrent tumour-specific chromosomal abnormalities, including rearrangements and copy number changes, has shown increasingly important diagnostic, therapeutic and prognostic implications. In surgical pathology, fluorescence in situ hybridization (FISH) is a quick and reliable method for the detection of many genetics aberrations, and can be applied to a variety of specimens including formalin-fixed paraffin-embedded (FFPE) tissues; as such, it has been successfully incorporated into diagnostic practice for lymphomas, mesenchymal and other solid tumours. The aim of this review is to discuss the utility, strengths and limitations of FISH when applied to the study of tumour tissue sections, with particular focus on the challenges associated with interpretation.

Harmonization of diagnostic terminology used during the histopathologic analysis of rodent tissue sections from nonclinical toxicity studies will improve the consistency of data sets produced by laboratories located around the world. The INHAND Project ( International Harmonization of Nomenclature and Diagnostic Criteria for Lesions in Rats and Mice) is a cooperative enterprise of 4 major societies of toxicologic pathology to develop a globally accepted standard vocabulary for proliferative and nonproliferative lesions in rodents. A prior manuscript ( Toxicol Pathol 2012;40[4 Suppl]:87S-157S) defined multiple diagnostic terms for toxicant-induced lesions, common spontaneous and age-related changes, and principal confounding artifacts in the rat and mouse central nervous system (CNS) and peripheral nervous system (PNS). The current article defines 9 new diagnostic terms and updates 2 previous terms for findings in the rodent CNS and PNS, the need for which has become evident in the years since the publication of the initial INHAND nomenclature for findings in rodent neural tissues. The nomenclature presented in this document is also available electronically on the Internet at the goRENI website ( http://www.goreni.org/ ).

The locomotor networks in the spinal cord coordinate muscle contractions and govern limb movements, ensuring postural body stability and smooth locomotion at different speeds. Dissecting the organization and connection of spinal motor networks is crucial to interpret their functional roles and thus can help to design more specific interventions for motor system diseases. Traditional two-dimensional (2D) histological sectioning is inadequate to accurately dissect complex architecture of the spinal cord as it provides only partial spatial information about spinal neural circuits. It is particularly difficult for quantitative analysis of axon regeneration after injury with 2D tissue sections, because it shows axonal fragments rather than spatial trajectory of regenerating axons. Therefore, three-dimensional (3D) imaging and analysis are extremely necessary for investigations of spinal cord structure and function. Although 3D spatial structure of the spinal cord can be reconstructed by serial sectioning, this approach is laborious and prone to image distortion. The recently emerging tissue clearing technique enables 3D imaging of the entire spinal cord at cellular resolution without tissue sectioning. The development of tissue clearing contributes to revealing the organization and function of spinal circuits and elucidating associated mechanisms underlying certain behaviours in health and disease. In this paper, we give an overview of the current clearing methods and introduce available labelling and imaging techniques as well as data processing software. Finally, we demonstrate the recent applications of tissue clearing in the spinal cord. Tissue clearing technique provides a novel tool for 3D imaging and quantification of the spinal cord, and benefits investigations of structure-function relationship of spinal networks. This review might help researchers to find the potential of tissue clearing in the studies of spinal cord and select appropriate clearing protocol for their experimental schemes.

The proper handling of the gross specimen is imperative, as it is the most important first step in providing excellent patient care. Our diagnoses depend on the correct description and submission of tissue sections for histologic analysis. A logical and problem-solving approach to handling the gross specimen is presented.

Objective Owing to variations in the exterior appearances of noncancerous diseases in the oral cavity, clinicians may have difficulty diagnosing oral squamous cell carcinoma (OSCC). Tissue biopsy is confirmatory, but invasive. Therefore, reliable tumor markers for OSCC are required. Here, exosomal Alix (exoAlix) levels were measured in serum/salivary samples from patients with OSCC and healthy controls (HCs). Methods Fifty-seven patients admitted to Nagoya University Hospital from 2017 through 2019 were enrolled, and serum samples (OSCC, n = 29; HC, n = 21) and/or saliva samples (OSCC, n = 23; HC, n = 20) were collected. Exosomal fractions were isolated using ultracentrifugation. ExoAlix levels were measured using enzyme-linked immunosorbent assay. Results Serum/salivary exoAlix levels were significantly higher in patients with OSCC than in HCs. Receiver operating characteristic analyses revealed that sensitivity, specificity, positive predictive value, and area under the curve were 0.345, 1.000, 1.000, and 0.685, respectively, for serum exoAlix and 0.348, 1.000, 1.000, and 0.712, respectively, for salivary exoAlix at optimal cut-off values (serum, 0.205; saliva, 0.193). All tested OSCC tissue sections (n = 21) were immuno-reactive for Alix. Conclusion Serum and salivary exoAlix were identified as potential diagnostic OSCC biomarkers. Serum exoAlix was suitable for prediction of therapeutic responses.

Objective The purpose of this study was to compare the acute histological effects of MicroPulse transscleral cyclophotocoagulation (MPCPC) using the MicroPulse P3 Device and continuous wave transscleral cyclophotocoagulation (CWCPC) on the ciliary body and adjacent structures in human cadaver eyes. Methods Quadrants of 6 human cadaver eyes from 3 different donors were subjected to traditional CWCPC, slow burn CWCPC, MPCPC, or no treatment (internal control). Sutures were used to differentiate different treatment areas on each eye. Differential inking was applied after treatments to aid in microscopic correlation. All specimens were subject to standard histologic processing. Tissue sections were cut at 4 microns and stained with hematoxylin and eosin according to established protocols. Pathologic evaluation by light microscopy was confirmed by a senior pathologist blinded to treatment groups. Results In all 6 eyes, tissues treated with traditional and low burn CWCPC showed variable coagulative tissue damage to the ciliary body compared with untreated tissues. Minimal histologic changes were identified within the ciliary processes, although variable pigment clumping and streaming were noted within the pigmented ciliary epithelium. In contrast to CWCPC, MPCPC-treated tissues showed only minimal coagulative tissue damage to the ciliary body. Variable pigment clumping and streaming, however, were also noted in the pigmented ciliary epithelium in MPCPC-treated tissues. Conclusions In human cadaver eyes, MPCPC treatment caused less tissue disruption to the ciliary body compared with traditional and low burn CWCPC treatments. MPCPC may be a less destructive and more selective method of cyclophotocoagulation when compared with traditional and low burn CWCPC.

Localized argyria is a benign rare condition resulting from impregnation of silver particles in the skin by various exposure sources such as silver jewelry,1 occupational exposures,2 topical medications,3 and medical procedures (ie, acupuncture).4 We report a case of acupuncture-induced argyria on the face, both dorsa of the hands, wrists, legs, and dorsa of the feet, successfully treated with 1 to 2 sessions of Q-switched 1064-nm neodymium-doped yttrium aluminum garnet (Nd:YAG) laser. Tissue section demonstrated refractile granules around the eccrine glands under dark-field microscopy.