The Application of Luminol-Bound Microspheres for the Quantitative Analysis of Toxic Oxygen within Phagosomes

Abstract

Hydrophilic microspheres, with a diameter of about 2 μm, composed of glyceryl methacrylate and methacrylic acid were prepared. The microspheres could be stably stored in dispersed state and demonstrated little non-specific adherance to cell membranes. Luminol-bound microspheres, prepared by covalently binding luminol to microspheres, were phagocytosed by phagocytic cells without opsonization. Luminol bound to the microspheres was oxidized by toxic oxygen within phagosomes resulting in the generation of chem iluminescence. The chemiluminescence elicited by luminol-bound micro spheres was an effective method of measuring the candidacidal activity of the cells. The difference in phagosomal toxic oxygen versus extracellular toxic oxy gen was examined.