Morphometrical Methodology in Quantification of Biological Tissue Components

• Main Author: Blomgren, Bo
• Format: Doctoral Thesis
• Language: English
• Published: Uppsala universitet, Institutionen för kvinnors och barns hälsa 2004
• Subjects: Bildanalys; Morphometry; Stereology; Image analysis; histology
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-4628; http://nbn-resolving.de/urn:isbn:91-554-6068-2

<b>Objective:</b> 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. <b>Material and methods:</b> 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. <b>Results:</b> The results show that computer-assisted morphometry is of great value for quantitative measurements of the following tissue components: <b>Epithelium:</b> The epithelial structure, instead of just thickness, was measured in an unbiased way. <b>Collagen:</b> The collagen fibril diameter was determined in electron microscopic specimens, and the collagen content was analysed in light microscopic specimens. <b>Elastic fibres:</b> The amount of elastic fibres in the connective tissue was measured after visualisation by autofluorescence. <b>Vasculature:</b> 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. <b>Smooth muscle:</b> 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. <b>Conclusion:</b> 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.