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01.08.2011 | Original Paper | Ausgabe 4/2011 Open Access

Cellular Oncology 4/2011

Rapid quantification of myocardial fibrosis: a new macro-based automated analysis

Cellular Oncology > Ausgabe 4/2011
Awal M. Hadi, Koen T. B. Mouchaers, Ingrid Schalij, Katrien Grunberg, Gerrit A. Meijer, Anton Vonk-Noordegraaf, Willem J. van der Laarse, Jeroen A. M. Beliën
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1007/​s13402-011-0035-7) contains supplementary material, which is available to authorized users.
This paper is a reprint from rapid quantification of myocardial fibrosis: a new macro-based automated analysis, Awal M. Hadi, Koen T.B. Mouchaers, Ingrid Schalij, Katrien Grunberg, Gerrit A. Meijer, Anton Vonk-Noordegraaf, Willem J. van der Laarse and Jeroen A.M. Belien, originally published in Analytical Cellular Pathology/Cellular Oncology, Volume 33, number 5–6, 2010, pp. 257–269, IOS Press.
An erratum to this article can be found at http://​dx.​doi.​org/​10.​1007/​s13402-011-0057-1



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.


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.


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.


Automated quantification of fibrosis is feasible, objective, and time-efficient.

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