Collagen IV and laminin‐1 expression in embryonic mouse lens using principal components analysis technique

Abstract

Immunohistochemistry section staining is not always easy to interpret. Manual quantification of immunohistochemical staining is limited by the observer visual ability to detect changes in level staining. Hence, the quantification of immunostaining by means of digital image analysis allows us to measure accurately protein expression percentages in immunobiological stained tissues and ensures to overcome the visual limitations. We perform an experimental study to analyse the impact of folic acid (FA) deficiency into collagen IV and laminin‐1 expression in the embryonic mouse lens. The study starts with microscope images of embryos mouse lens whose mothers fed a diet deficient in FA during 2 and 8 weeks. A principal component analysis (PCA) image processing is used to analyse these images coming from control and FA deficit groups. The method permits to define an index of over‐ or infraexpression of collagen IV and laminin‐1 associated to different spatial organisation structures (PC processes). Additionally, it permits to determine in precise percentage the exact quantity of the overexpression or infraexpression and finally to comprehend molecular regionalisation and expression in both control and deficient groups. The results suggest that even with 2 weeks of deficit of FA the expression and distribution of both molecules is affected.