Attenuator to suppress extraneous light in luminescent specific-binding

Abstract

An attenuator is included in a reagent medium of a luminescent specific-binding assay to suppress undesirable extraneous light. In one such assay, an analyte in a sample is reacted with a specific binding partner attached to a solid surface, forming an immobilized pair at the surface. One member of the immobilized pair is then allowed to react with a specific binding partner previously conjugated with one component of a luminescent reaction system, and the remaining components are provided in the reagent medium. The resulting light emitted in the luminescent reaction is recorded on photographic film or other photodetector as a measure of the presence and quantity of the analyte in the sample. The reagent medium containing the remaining luminescent reaction system components also includes an attenuator, preferably a dye, which absorbs light over a range of wavelengths including that of the light emitted in the luminescent reaction, the attenuator being present in an amount sufficient to suppress the extraneous light observed adjacent the solid surface in the absence of the attenuator. Reduction of such extraneous light sharpens the recorded luminescent image, thereby allowing a more precise analysis of the light intensity, and additionally reduces the occurrence of false positive images in the assay. In another embodiment, the attenuator preferentially absorbs light emitted in remote portions of a reaction volume so that only light emitted from proximal portions reaches a measurement means. Reactions otherwise requiring a separation step may thereby be conducted homogeneously.