Miniaturization of luminometric assays: Optimizing the performance of common luminescent assays in 384-, and 1536-well plate formats

Jorma Lampinen, Marika Raitio and Reija-Riitta Harinen, Microplate Instrumentation, Thermo Fisher Scientific, Vantaa, Finland

Increasing use of small assay volumes with high density plates is a clear trend in today's bioscience research. Most of the commercial assay kits are still designed for 96 well plates and all instructions are given for that plate format. Therefore, the user needs most often to modify the assay procedure to be able to use low volume/high density microplates. There are no general rules how this modification is done making miniaturization of the assays sometimes complicated. In this paper we have optimized the performance or common luminometric assays for 384, 384 low volume and 1536 well plates. The key parameters to be followed to evaluate the assay performance with the high density plate assays were the detection limit (LOD), CV%, and Z-prime. Based on these results, it can be recommended that, considering all aspects (reagent and material consumption, performance etc.), the optimal assay setup would be 384-low volume plate with about 10 total assay volumes.