Automated slide scanning and segmentation of fluorescently-labeled tissues is the most efficient way to analyze whole slides or large tissue sections. Unfortunately, many researchers spend large amounts of time and resources developing and optimizing workflows that are only relevant to their own experiments. In this article, we describe a protocol that can be used by those with access to a widefield high-content analysis system (WHCAS) to image any slide-mounted tissue, with options for customization within pre-built modules found in the associated software. Not originally intended for slide scanning, the steps detailed in this article make it possible to acquire slide scanning images in the WHCAS which can be imported into the associated software. In this example, the automated segmentation of brain tumor slides is demonstrated, but the automated segmentation of any fluorescently-labeled nuclear or cytoplasmic marker is possible. Furthermore, there are a variety of other quantitative software modules including assays for protein localization/translocation, cellular proliferation/viability/apoptosis, and angiogenesis that can be run. This technique will save researchers time and effort and create an automated protocol for slide analysis.
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http://dx.doi.org/10.3791/57440 | DOI Listing |
Toxicol Pathol
December 2024
AbbVie Inc., North Chicago, Illinois, USA.
Enhanced histopathology of the immune system uses a precise, compartment-specific, and semi-quantitative evaluation of lymphoid organs in toxicology studies. The assessment of lymphocyte populations in tissues is subject to sampling variability and limited distinctive cytologic features of lymphocyte subpopulations as seen with hematoxylin and eosin (H&E) staining. Although immunohistochemistry is necessary for definitive characterization of T- and B-cell compartments, routine toxicologic assessments are based solely on H&E slides.
View Article and Find Full Text PDFJ Pathol Inform
January 2025
U.S. Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Imaging, Diagnostics, and Software Reliability, Silver Spring, MD, United States of America.
Objective: With the increasing energy surrounding the development of artificial intelligence and machine learning (AI/ML) models, the use of the same external validation dataset by various developers allows for a direct comparison of model performance. Through our High Throughput Truthing project, we are creating a validation dataset for AI/ML models trained in the assessment of stromal tumor-infiltrating lymphocytes (sTILs) in triple negative breast cancer (TNBC).
Materials And Methods: We obtained clinical metadata for hematoxylin and eosin-stained glass slides and corresponding scanned whole slide images (WSIs) of TNBC core biopsies from two US academic medical centers.
J Pathol Inform
December 2024
Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Lisbon 1049-001, Portugal.
Whole slide images (WSI), obtained by high-resolution digital scanning of microscope slides at multiple scales, are the cornerstone of modern Digital Pathology. However, they represent a particular challenge to artificial intelligence (AI)-based/AI-mediated analysis because pathology labeling is typically done at slide-level, instead of tile-level. It is not just that medical diagnostics is recorded at the specimen level, the detection of oncogene mutation is also experimentally obtained, and recorded by initiatives like The Cancer Genome Atlas (TCGA), at the slide level.
View Article and Find Full Text PDFSud Med Ekspert
December 2024
Russian Center of Forensic Medical Expertise, Moscow, Russia.
Objective: To examine the changes in dermal mast cells density in the mechanically injured skin and to evaluate the applicability of dermal mast cells density increase as an injuries vitality diagnostic criteria (serial examination, statistically processed results).
Material And Methods: 240 skin autopsy samples with mechanical injuries from 40 persons were divided to 3 groups (80 in each group): vital injuries, postmortal injuries, control non-injured samples. A routine histological examination using standard H&E stain and IHC with mast cells tryptase antibodies was performed consequented with histological slides full-format scanning and computer processing.
Int J Surg Case Rep
December 2024
Resident of Orthopaedics and Traumatology Department, Faculty of Medicine, Universitas Brawijaya - Saiful Anwar General Hospital, Malang, East Java, Indonesia.
Introduction: Hip fractures are common in elderly patients, often accompanied by comorbidities. These fractures can be combined with other injuries, such as a femoral head, neck, or shaft dislocation. The cause of this complex injury is not well-established, but factors like high-energy trauma or falls from a certain height may contribute.
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