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Advances in Translational Pathology

Brightfield microscopy is the preferred method of pathologists for diagnosing solid tumors, utilizing common staining techniques such as hematoxylin and eosin staining and immunohistochemistry. The editors of Laboratory Investigation present a collection of articles that describe new ways to process samples and interpret data generated by histopathology and immunohistochemistry, as well as new technologies that complement these methods.

Histopathology

Polarization-second harmonic microscopy was utilized to investigate whether collagen ultrastructure in thyroid due to four carcinoma types and Graves’ disease could be differentiated in human histopathology samples. Three parameters were extracted, revealing that the degree of linear polarization and χ(2)zzz/χ(2)zxx were effective in differentiating some diseases, while the parameter χ(2)xyz/χ(2)zxx was less effective.

Article | | Laboratory Investigation

Accurate quantification of steatosis in liver biopsies is a key step in the treatment of patients with fatty liver diseases. To assist pathologists for such analysis tasks, we develop a novel deep learning-based framework to segment overlapped steatosis droplets in whole slide liver biopsy images. Quantitative measurements of steatosis at both pixel and object-level present strong correlation with clinical data, suggesting its potential for clinical decision support.

Technical Report | | Laboratory Investigation

Real-time lipid patterns can identify liver tumors and their inter-tumor and intra-tumor heterogeneity. Ceramides and related sphingolipids are a common feature of necrotic tumors and can characterize the tumor phenotype based on metabolic shifts relevant for cell death pathways. Lipid patterns have the potential to improve clinical decision-making in the near future.

Article | | Laboratory Investigation

Mucus was tested as a potential biological material for screening/early diagnosis of colorectal cancer using infrared spectroscopy. Based on a digital histopathology and statistical modeling approach, cancerous and non-cancerous samples were classified with an area under the curve performance of 95% based on mucus spectral profiles, indicating changes in the glycan component of mucins.

Article | Open Access | | Laboratory Investigation

The paper describes a fully automated numerical analysis for an unbiased quantification of non-alcoholic fatty liver disease (NAFLD) histological features in rodent models. The technique offers a quantitative high-throughput method to rapidly detect NAFLD in large preclinical studies and for accurately monitoring disease evolution.

Technical Report | | Laboratory Investigation

In this paper, the authors describe the development and validation of a novel image signature-based radiomics model. A total of 655 glioma patients were enrolled to build this model which is shown to be an effective tool to achieve multilayer preoperative diagnosis and prognostic stratification of gliomas.

Article | | Laboratory Investigation

The NanoSuit method permits the study of paraffin sections using correlative light and electron microscopy at low and high magnification, with the following features: (i) the integrity of the glass slide is maintained, (ii) 3D microstructures of tissue and pathogens can be visualized, (iii) nuclei and 3,3′-diaminobenzidine-stained areas are distinct because of gold chloride usage, (iv) immunohistochemical staining is quantitative, and (v) contained elements can be analyzed.

Technical Report | Open Access | | Laboratory Investigation

Immunohistochemistry

Proteomic profiling may contribute to the analysis and classification of cancer. The authors applied the digital western blot technique DigiWest with a panel of 102 proteins and phosphoproteins in combination with a machine learning algorithm to classify the tissue origin of five common cancer types in fresh frozen and formalin-fixed paraffin-embedded tissue. DigiWest profiling represents a valuable method for cancer classification, yielding conclusive and decisive data, thus making this approach attractive for routine clinical applications.

Article | Open Access | | Laboratory Investigation

The authors developed a high-content, quantitative analysis of breast cancer tissues based on microfluidic staining and image processing, to characterize both HER2 overexpression and amplification at the cellular level. This study paves the way for evaluatation of intratumoral heterogeneity with unprecedented accuracy with standard staining methods such as immunofluorescence and FISH.

Technical Report | | Laboratory Investigation

The authors developed a novel simplified assay for glioblastoma transcriptional classification on formalin-fixed-paraffin-embedded tissue samples. On such dataset, immunohistochemical profiles, based on expression of a restricted panel of gene classifiers, were integrated by machine learning approach to generate a glioblastoma transcriptional signature based on protein quantification that allowed to efficiently assign transcriptional subgroups to an extended cohort. Correlations with both histopathological features and clinical outcome have been also performed.

Article | | Laboratory Investigation

Digital image analysis (DIA) of multiplex fluorescence-based immunohistochemistry and visual chromogenic evaluation of CDX2, SOX2, SOX9, E-cadherin, and β-catenin in colorectal cancer are comparable, recognizing prognostic value of CDX2 and negative correlation with SOX2. Membrane staining is best evaluated visually, while DIA enables single-cell coexpression analysis and improves visualization and detection of clinicopathological and biological associations.

Technical Report | Open Access | | Laboratory Investigation

Digital spatial profiling is a new high-plex technology with potential to multiplex hundreds of proteins on a single slide. Here the authors validate the digital aspect of the technology on a control tissue microarray with known amounts of PD-L1 expression to show it has quantitative capacity comparable to quantitative immunofluorescence.

Article | | Laboratory Investigation

PD-L1 IHC was evaluated in stored tissue with mass spectrometry (MS). Increased humidity and temperature resulted in considerable immunoreactivity loss, particularly among antibodies recognizing extracellular and discontinuous epitopes, which could be partially mitigated with the use of desiccant. However, even in significantly affected tissues MS was able to quantitate PD-L1 expression and assess peptide oxidation. These results suggest MS is suited for biomarker assessments using stored sections—a sample type commonly encountered in oncology research.

Article | Open Access | | Laboratory Investigation

Detailed protocols for immunohistochemical and in situ hybridization assays for the detection of SARS-CoV-2 are provide so they can be readily implemented in pathology laboratories and medical examiner offices for diagnostic and research purposes. These assays were found to represent a sensitive and specific method for detecting the virus in tissue samples.

Technical Report | | Laboratory Investigation