Morphometrical Methodology in Quantification of Biological Tissue Components

Sammanfattning: Objective:To develop and validate computer-assisted morphometrical methods, based on stereological theory, in order to facilitate the analysis and quantitative measurements of biological tissue components.Material and methods:Biopsy specimens from the vaginal wall or from the vestibulum vaginae of healthy women, or from women suffering from incontinence or vestibulitis were used.A number of histochemical methods for light microscopy were used, and modified for the different morphometrical analyses. Electron microscopy was used to reveal collagen fibre diameter.Computer-assisted morphometry, based on image analysis and stereology, was employed to analyse the different tissue components in the biopsies. Computer programs for these purposes were developed and validated.Results:The results show that computer-assisted morphometry is of great value for quantitative measurements of the following tissue components:Epithelium: The epithelial structure, instead of just thickness, was measured in an unbiased way.Collagen: The collagen fibril diameter was determined in electron microscopic specimens, and the collagen content was analysed in light microscopic specimens.Elastic fibres: The amount of elastic fibres in the connective tissue was measured after visualisation by autofluorescence.Vasculature: A stereological method using a cycloid grid was implemented in a computer program. Healthy subjects were compared with patients suffering from vestibulitis. The results were identical in the two groups.Smooth muscle: A stereological method using a point grid was implemented in a computer program. Using the Delesse principle, the fibres were calculated as area fractions. The area fractions were highly variable among the different specimens.Conclusion:Morphometry, used correctly, is an important analysis method in histopathological research. It is important that the methods are as simple and user-friendly as possible. The present studies show that this methodology can be applied for most quantitative histological analyses.