Chapter 6: Understanding Luminescence Based Screens

Luminescent assays represent a large proportion of the assays found in high throughput screening applications. The widespread use of these assays is due to the desirable high sensitivity of the luminescent signal. Luminescence can arise from enzymatic or chemical reactions and in this chapter we provide a description of some of the most common bioluminescent and chemiluminescent assay formats. A description of luciferase enzymes, which are often found in cell based assays, is given along with recommendations for the optimal use of assays employing these enzymes. One of the most common chemiluminescent assays based on luminescent oxygen channeling is also described, along with considerations for the design and implementation of this assay format. Optimal use of these assays requires an appreciation of luminescent light generation mechanisms and the interferences that can occur when applying these to large collections of compounds. Orthogonal and counter screen approaches are outlined in this chapter, both of which are necessary to understand luminescence based screens.