In Situ Microdensitometry of Neoglycoprotein Staining: Detection of Tissue-Site Specific Patterns of Endogenous Lectin Expression
In this chapter we introduce a microdensitometric and image analysis procedure for the in situ quantification of endogenous lectin expression by normal and altered cells in different tissues and even at specific sites within a single tissue. The large family of sugar-binding molecules can be detected by biotinylated neoglycoproteins (see Danguy an Gabius, Chap. 25, and Sinowatz et al, Chap. 27, this vol.), using a peroxidase reaction associated to the avidin-biotin interaction. The chromophore concentration precipitated at defined tissue sites and components is proportional to the number of sugar binding sites, and quantification can be done by microdensitometry, i.e., by registfating the optical density (light intensity in negative images) at specific tissue areas (pixels) of digitalized images.
KeywordsSugar Hydrogen Peroxide Albumin Carbohydrate Heparin
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