Analysis of CCN Expression by immunofluorescence on skin cells, skin, and reconstructed epidermis

Abstract

During a long time, immunofluorescence has been neglected to benefit of molecular biology especially genetics, transcriptomics, and proteomics analyses. These techniques give good results on cell culture but for organs that are made of numerous cells with several compartments, various states of differentiation as in epidermis, immunohistochemistry is always relevant. Double (triple) staining by immunofluorescence allows positive cells identification in complex cell structure (for example, pericytes and endothelial cells in vessels) and subcellular localizations. In order to, due to improvement of antibodies avoiding especially species cross-reactions, microscopy and specific softwares, quality of staining, and acquired images have been upgraded. Consequently, this technique permits, as molecular biology analyses, quantification of the level of expression as intensity of fluorescence can be measured in each cells and each compartments (nuclear, cytoplasmic). In order to immunofluorescence on cells and tissue needs few materials and gives at the same times qualitative and quantitative results and must be used more widely especially when a mutation was associated to a disease.