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Liver fibrosis is the excessive accumulation of extracellular matrix proteins including collagen that occurs in most types of chronic liver diseases. Research into novel anti-fibrotic therapies is required to meet important patient needs in this area.
Collagen and alpha Smooth Muscle Actin (αSMA) content can be used to reveal fibrotic changes within the liver. Whole slide liver sections can be analysed to classify and quantify established lesional (fibrotic) areas and normal parenchyma as regions of interest (ROI). Within these stained ROI, the amount of collagen (highlighted with Picrosirius red histological stain) and αSMA content (highlighted by αSMA IHC) can be accurately quantified and the data can be used to determine the effect of drug treatment on liver fibrosis.
A custom algorithm is developed to accurately detect collagen within the PSR stained sections and αSMA positive staining within the αSMA stained sections across the liver tissue ROI. Stain intensity thresholds within each algorithm are established per marker to classify positive staining as low (yellow overlay mask), medium (orange overlay mask) and high (red overlay mask). Once developed, algorithms are objectively applied to each set of images to quantify and classify the area of positive staining.
Example Readouts Include:
Area of detected liver tissue ROI per whole section in mm2
Area of Collagen (based on PSR threshold level) within the liver tissue ROI per slide.
Classification of detected PSR area stain intensity as low, medium and high.
Area of αSMA stain (based on DAB threshold level) within the liver tissue ROI per slide.
Classification of detected αSMA stain area intensity as low, medium and high.
% collagen and % αSMA within the whole tissue ROI per slide.
Example PSR Analysis & Data
Example PSR Analysis. A) Original histologically stained PSR whole image. B) Classified regions in whole image showing: Liver tissue (red overlay), Artefact (green overlay), White space (white overlay). C) Collagen detection in PSR whole image classified as: Low intensity (yellow overlay), Moderate intensity (orange overlay), High intensity (red overlay). D) Original histologically stained PSR image at high magnification from box area. E) Classified regions at high magnification. F) Collagen detection in PSR image at high magnification.
PSR Liver Fibrosis Analysis Data
Example αSMA Analysis & Data
Example αSMA Analysis. A)Original αSMA IHC whole image. B)Classified regions in whole image showing: Liver tissue (red overlay), Artefact (green overlay) White space (white overlay). C) αSMA Positive Area detection in whole image classified as: Low intensity (yellow overlay), Moderate intensity (orange overlay), High intensity (red overlay). D) Original αSMA IHC at high magnification from box area. E) Classified regions at high magnification. F) αSMA positive area detection at high magnification.
αSMA Analysis Data
Custom algorithms allow accurate classification of regions of interest (ROI)
Accurate segmentation of tissue from white space areas across whole section
The ability to quantify and classify the area of collagen staining within each ROI
The ability to quantify and classify the area of αSMA staining within each ROI
Objectively applied analysis across all study slides
Whole slide quantitative analysis increases the robustness of the data
Whole slide image overlays of ROI, collagen and αSMA detection returned to the Client