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Updated: Jul 15, 2025
Author Spotlight: Advancing Human Cardiac Anatomy Through Multi-Scale Analysis of Hearts
Published on: June 28, 2024
Semiautomated pipeline for quantitative analysis of heart histopathology
Patrick Droste1,2, Dickson W L Wong1, Mathias Hohl3
1LaBooratory of Nephropathology, Institute of Pathology, Medical Faculty, RWTH Aachen University, Aachen, Germany.
Insights
A new ImageJ macro, HeartJ, offers rapid, automated quantification of cardiomyocyte size and capillary density in heart tissue. This tool aids research into heart disease by providing objective histological analysis for both animal models and human samples.
Area of Science:
- Cardiovascular Research
- Histology and Pathology
- Medical Imaging Analysis
Background:
- Heart diseases cause significant global mortality, often leading to pathological cardiomyocyte hypertrophy and capillary rarefaction.
- Quantifying these cardiac changes in patients and animal models is technically challenging and time-consuming.
- Existing methods for assessing cardiac hypertrophy and microvasculature density are often laborious and slow.
Purpose of the Study:
- To develop a semiautomated pipeline for quantifying cardiomyocyte size and capillary density in cardiac histology.
- To provide a rapid, objective, and accessible tool for researchers studying heart disease.
- To validate the pipeline's translatability between animal models and human clinical samples.
Main Methods:
- Development of ImageJ macros, termed HeartJ, for semiautomated histological analysis.
- Utilized modified Gomori silver staining and Fluorescein-labeled lectin staining for capillary visualization.
- Validated the HeartJ pipeline in a mouse model of cardiac hypertrophy and human autopsy samples.
Main Results:
- HeartJ enabled accurate quantification of cardiomyocyte hypertrophy and microvasculature rarefaction in both murine and human cardiac samples.
- Confirmed similar cardiomyocyte diameters and hypertrophy extent between mice and humans, validating the animal model.
- Demonstrated the pipeline's ability to analyze protein expression in individual cardiomyocytes.
Conclusions:
- The HeartJ pipeline facilitates effective and objective quantitative histological analyses in preclinical and clinical heart samples.
- This tool significantly speeds up analysis, making complex histological assessments feasible.
- HeartJ is freely available, has minimal hardware requirements, and supports diverse staining techniques for comprehensive cardiac research.
Background:
Heart diseases are among the leading causes of death worldwide, many of which lead to pathological cardiomyocyte hypertrophy and capillary rarefaction in both patients and animal models, the quantification of which is both technically challenging and highly time-consuming. Here we developed a semiautomated pipeline for quantification of the size of cardiomyocytes and capillary density in cardiac histology, termed HeartJ, by generating macros in ImageJ, a broadly used, open-source, Java-based software.
Methods:
We have used modified Gomori silver staining, which is easy to perform and digitize in high throughput, or Fluorescein-labeled lectin staining. The latter can be easily combined with other stainings, allowing additional quantitative analysis on the same section, e.g., the size of cardiomyocyte nuclei, capillary density, or single-cardiomyocyte protein expression. We validated the pipeline in a mouse model of cardiac hypertrophy induced by transverse aortic constriction, and in autopsy samples of patients with and without aortic stenosis.
Results:
In both animals and humans, HeartJ-based histology quantification revealed significant hypertrophy of cardiomyocytes reflecting other parameters of hypertrophy and rarefaction of microvasculature and enabling the analysis of protein expression in individual cardiomyocytes. The analysis also revealed that murine and human cardiomyocytes had similar diameters in health and extent of hypertrophy in disease confirming the translatability of our murine cardiac hypertrophy model. HeartJ enables a rapid analysis that would not be feasible by manual methods. The pipeline has little hardware requirements and is freely available.
Conclusions:
In summary, our analysis pipeline can facilitate effective and objective quantitative histological analyses in preclinical and clinical heart samples.
