Many current microarray technologies rely on immobilization of ligands using a single crosslinking method. Here the effects of various crosslinkers on binding of cholera toxin and other biological targets to immobilized carbohydrates were assessed. With each of the linkers used, the chemical moieties targeted for crosslinking (amines, thiols) were identical, but the physical and chemical properties of the tethers were varied. Although antibody binding was largely unaffected by tether characteristics, binding of toxin to immobilized carbohydrates was greatly affected by the nature of the tether, indicating that a '' one-size-fits-all '' approach to array production may not be appropriate in many cases.