E-Book Overview
A comprehensive collection of optimized methods for using lectins in biomedical research. The techniques-fine-tuned by the researchers who developed them-apply lectins to the analysis of carbohydrates, to the isolation and quantification of glycoproteins, and to living cells. Also included are detailed methods for the separation of human bone marrow cells for clinical use, for the assessment of the biological effects of plant lectins, and for the use of light and electron microscopy in analyzing carbohydrate structures. Extremely useful tools for examining structural changes in glycosylation and their functional consequences for human pathophysiology.
E-Book Content
1 Light Microscopy Overview and Basic Methods Anthony
J. Leathem and Susan A. Brooks
1. Introduction 1.1. Why Use Lectins? Lectms are proteins or glycoprotems of nonimmune origin derived from plants, animals, or mlcroorgamsms that have specn7cIty for termmal or subterminal carbohydrate residues. They are sensitive, stable, and easy-to-use tools. Lectm hlstochemlstry and cytochemistry can provide an extraordmarlly sensettve detection system for changes m glycosylatlon and carbohydrate expression that may occur during embryogenesis, growth, and disease. Although carbohydrates occur in a vast range of permutations that may be present m and between cells, there 1sfrequently a dominance and conservation of structures to give specific markers of cell types or differentiation. Lectm hlstochemlstry or cytochemistry can reveal subtle alterations in glycosylatlon between otherwise mdistmgulshable cells. Lectins wtll specifically recogmze and bmd to carbohydrate structures on the surface of cells, on cytoplasmic and nuclear structures, and m extracellular matrix m cells and tissues from throughout the ammal and plant kingdoms, down to bacteria and vu-uses. 7.2. What Do Lectins Bind to? Sugar combmmg speci