Key result
Semi-automated macro-based image analysis for quantifying myocardial fibrosis correlated strongly with stereology (r2=0.95) and required only 22% of the time needed for traditional methods.
Why the study?
Does a semi-automated ImageJ macro accurately and efficiently quantify myocardial fibrosis compared to traditional methods in rat and human tissues?
Does a semi-automated ImageJ macro accurately and efficiently quantify myocardial fibrosis compared to traditional methods in rat and human tissues?
Effect estimate: r2=0.95
A new semi-automated macro for ImageJ provides a rapid, objective, and accurate method for quantifying myocardial fibrosis compared to traditional stereology and polarisation microscopy.
Facilitates rapid fibrosis quantification in preclinical models; leaves open validation for clinical workflows.
Background: Fibrosis is associated with various cardiac pathologies and dysfunction. Current quantification methods are time-consuming and laborious. We describe a semi-automated quantification technique for myocardial fibrosis and validated this using traditional methods. Methods: Pulmonary Hypertension (PH) was induced in adult Wistar rats by subcutaneous monocrotaline (MCT) injection (40 mg/kg). Cryosections of myocardial tissue (5 μm) of PH rats (n=9) and controls (n=9) were stained using Picrosirius red and scanned with a digital microscopic Mirax slide scanner. From these sections 21 images were taken randomly of each heart. Using ImageJ software a macro for automated image analysis of the amount of fibrosis was developed. For comparison, fibrosis was quantified using traditional polarisation microscopy. Both methods were correlated and validated against stereology as the gold standard. Furthermore, the method was tested in paraffin-embedded human tissues. Results: Automated analysis showed a significant increase of fibrosis in PH hearts vs. control. Automated analysis correlated with traditional polarisation and stereology analysis (r2=0.92 and r2=0.95, respectively). In human heart, lungs, kidney and liver, a similar correlation with stereology (r2=0.91) was observed. Time required for automated analysis was 22 and 33% of the time needed for stereology and polarisation analysis, respectively. Conclusion: Automated quantification of fibrosis is feasible, objective and time-efficient.
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Mouchaers et al. (2010) studied Myocardial Fibrosis (n=18). Semi-automated macro-based image analysis vs. Traditional polarisation microscopy and stereology was evaluated on Correlation with traditional polarisation and stereology analysis (r2=0.95). Semi-automated macro-based image analysis for quantifying myocardial fibrosis correlated strongly with stereology (r2=0.95) and required only 22% of the time needed for traditional methods.
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