Rapid Quantification of Myocardial Fibrosis: A New Macro-Based Automated Analysis

Abstract

Abstract.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 (r 2 = 0.92 and r 2 = 0.95, respectively). In human heart, lungs, kidney and liver, a similar correlation with stereology (r 2 = 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.

Development of a Computational High-Throughput Tool for the Quantitative Examination of Dose-Dependent Histological Features

“…Comparative analysis indicates QuHAnT accurately quantitated increases in lipid vacuolization compared to manual point counting as demonstrated by the high V v value concordance between these approaches. These results confirm that QuHAnT is a valuable complement to pathologist-directed assessments and suggest that it is a more comprehensive approach for the quantitation of histological features compared to manual point counting (Hadi et al 2010; Levene et al 2012; Garrido et al 2013). …”

“…Consequently, the manual quantitation of collagen deposition would require a more comprehensive and labor-intensive assessment. For example, PSR quantitation is highly correlated to automated analysis when using a 200-point grid for manual quantitation (Hadi et al 2010). However, using a 200-point grid density or alternative manual approach is not practical when examining thousands of images further emphasizing the value of a high-throughput image analysis tool.…”

Persistent infection with Crohn’s disease-associated adherent-invasive Escherichia coli leads to chronic inflammation and intestinal fibrosis

“…Multiple sections from the same tissue from several animals at each time point were scored according to criteria and descriptions previously defined 43 and summarized in Supplementary Table S1. In some experiments, quantitative analysis of the PSR-stained images taken under polarized light was performed using an ImageJ macro-based algorithm as previously described 44 . Immunohistochemical (IHC) staining of formalin-fixed tissue sections was performed using antibodies against F4/80 (1:600) and CD3 (1:1000) at Histology Services (McMaster University) and FoxP3+ (1:XXX) and O83+ (1:500) staining was done at Vertex Pharmaceuticals (Cambridge MA).…”

Characteristics of Interstitial Fibrosis and Inflammatory Cell Infiltration in Right Ventricles of Systemic Sclerosis-Associated Pulmonary Arterial Hypertension

“…Care was taken not to include vessels into the selected areas to ensure only interstitial myocardial fibrosis was quantified. Interstitial myocardial fibrosis was assessed in the RV and LV, using a fully automated analysis according to Mouchaers [17]. …”