Analysis of Uterine Morphology in Ovariectomized Rats Treated With Alendronate and Hop Extract Using Open-Source Software


  • Edi Rođak Department for Histology and Embryology, Faculty of Medicine Osijek, University of Osijek, J. Huttlera 4, 31000 Osijek, Croatia
  • Nada Oršolić Department of Animal Physiology, Faculty of Science Zagreb, University of Zagreb, Rooseveltov trg 6, Zagreb, Croatia
  • Robert Grgac Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branišovská 1160/31, 37005 České Budějovice, Czechia Faculty of Science, University of South Bohemia, Branišovská 1645/31a, 37005 České Budějovice, Czechia
  • Jasmina Rajc Department of Pathology and Forensic Medicine, University Hospital Centre Osijek, J. Huttlera 4, 31000 Osijek, Croatia Department of Pathology and Forensic Medicine, Faculty of Medicine Osijek, University of Osijek, J. Huttlera 4, 31000 Osijek, Croatia
  • Marina Bakula Department of Pathology and Forensic Medicine, University Hospital Centre Osijek, J. Huttlera 4, 31000 Osijek, Croatia
  • Nikola Bijelić Department for Histology and Embryology, Faculty of Medicine Osijek, University of Osijek, J. Huttlera 4, 31000 Osijek, Croatia



alendronate, hop extract, image analysis, open-source, osteoporosis, uterus


Free and open-source software for image analysis and morphological measurements in scientific research is rising in popularity and capabilities as new methods, plugins and macros are being actively developed. A semi-automated method for measuring rat uterus morphology using free and open-source software (Gimp and FIJI) is demonstrated in this paper. Research was performed on ovariectomized rats as a model of osteoporosis (with sham-operated control group). The animals were treated with alendronate, hop extract or the combination of the two. Whole histological slides were photographed and images were manually pre-processed in Gimp. Color masks from Gimp were loaded in FIJI and polar transformation and measurements were made using a custom macro. This analysis was supplemented by manual assessment of Ki67 proliferation marker expression by a pathologist. Our results suggest that monotherapy or combination therapy with alendronate and hop extract does not cause proliferation of the endometrium in ovariectomized rats and would be safe for use in osteoporosis treatment in this regard. The semi-automated method used in this research is more precise and unbiased than older manual methods. Furthermore, it can be easily adapted for analysis of whole-slide images of almost any round or oval organ.


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How to Cite

Rođak, E., Oršolić, N., Grgac, R., Rajc, J., Bakula, M., & Bijelić, N. (2022). Analysis of Uterine Morphology in Ovariectomized Rats Treated With Alendronate and Hop Extract Using Open-Source Software. Image Analysis and Stereology, 41(3), 193–202.



Original Research Paper