Abstract
Infertility is increasing worldwide; male factors can be identified in nearly half of all infertile couples. Histopathologic evaluation of testicular tissue can provide valuable information about infertility; however, several different evaluation methods and semi-quantitative score systems exist. Our goal was to describe a new, accurate and easy-to-use quantitative computer-based histomorphometric-mathematical image analysis methodology for the analysis of testicular tissue. On digitized, original hematoxylin-eosin (HE)-stained slides (scanned by slide-scanner), quantitatively describable characteristics such as area, perimeter and diameter of testis cross-sections and of individual tubules were measured with the help of continuous magnification. Immunohistochemically (IHC)-stained slides were digitized with a microscope-coupled camera, and IHC-staining intensity measurements on digitized images were also taken. Suggested methods are presented with mathematical equations, step-by-step detailed characterization and representative images are given. Our novel quantitative histomorphometric-mathematical image analysis method can improve the reproducibility, objectivity, quality and comparability of andrological-reproductive medicine research by recognizing even the mild impairments of the testicular structure expressed numerically, which might not be detected with the present semi-quantitative score systems. The technique is apt to be subjected to further automation with machine learning and artificial intelligence and can be named ‘Computer-Assisted or -Aided Testis Histology’ (CATHI).
Highlights
IntroductionInfertility incidence and prevalence are increasing worldwide, up to 15% of reproductiveaged couples are affected by infertility according to the World Health Organization [1]
Infertility incidence and prevalence are increasing worldwide, up to 15% of reproductiveaged couples are affected by infertility according to the World Health Organization [1].According to the International Glossary on Infertility and Fertility Care, infertility can be defined as: ‘a disease characterized by the failure to establish a clinical pregnancy after 12 months of regular, unprotected sexual intercourse or due to an impairment of a person’s capacity to reproduce either as an individual or with his/her partner
Our novel quantitative histomorphometric-mathematical image analysis method can improve the reproducibility, objectivity, quality and comparability of andrological-reproductive medicine research by recognizing even the mild impairments of the testicular structure expressed numerically, which might not be detected with the present semi-quantitative score systems
Summary
Infertility incidence and prevalence are increasing worldwide, up to 15% of reproductiveaged couples are affected by infertility according to the World Health Organization [1]. The Bergmann-Kliesch score compares the number of tubules containing elongated late spermatids and sperm to the total number of tubules on the sample and gives 0–10 points according to the calculated percentages [6,7] These score systems are semi-quantitative and do not provide exact quantitative numerical details of the structure of the spermatogenic epithelium, tubules or testis tissue. The improvement of microscope and computer techniques made it possible to create a microscope-coupled camera, and several image analysis programs have been developed which allowed the examination of the live microscopic images Capture of these live microscopic images on fixed magnifications (5×, 10×, 20×, 40×, 100× or higher) resulted in digitized images where several various quantitative measurements on testicular components could be taken for further research purposes [41,42,43,44,45,46]. As basic and translational andrological researches are often carried out on rodents nowadays, we used rodent testicular tissue from one of our previous experiments; our technique could be applied to human testicular tissue
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