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Synergy™ HT: A Multi-Mode Microplate Reader for Today's High Performance Luminescence ResearchDownload
Related Products: Synergy HTX
June 14, 2005
Author: Paul Held, BioTek Instruments, Winooski, Vermont
Luminescent assays are sensitive and quantitative tools commonly used for a variety of purposes in biomedical and pharmaceutical research. Quantitation of ATP can be used to detect and measure cellular growth, as well as bacterial contamination. Luminescent genetic reporting assays are widely used to study gene expression and cellular responses to external stimuli in Prokaryotic and Eukaryotic organisms. Dual-reporter assays use two independent reporter systems simultaneously to improve experimental accuracy. One reporter is used to measure the response resulting from the experimental conditions and is often referred to as the “experimental” reporter. The second reporter is designed not to respond to the experimental conditions, acting as an internal control from which data generated by the experimental reporter can be normalized. Normalization of the data serves to compensate for variability caused by differences in transfection efficiency, cell viability, cell lysis, and pipetting. Promega’s Dual-Luciferase® System uses the activities of luminescent proteins (luciferases) from the firefly (Photinus pyralis) beetle and the sea pansy (Renilla renformis) to serve as an experimental and a control reporter respectively (See Figure 1).
The Synergy™ HT Multi-Detection Reader (BioTek Instruments, Winooski, VT) is a robotic compatible microplate reader that can measure absorbance, fluorescence, and luminescence in all plate formats up to 384-well plates with exceptional performance in all three detection modes, that has recently been optimized for luminescence measurements (Figure 2). The Synergy HT utilizes a unique dual optics design that has both a monochromator/xenon flash system with a silicone diode detector for absorbance and a tungsten halogen lamp with blocking interference filters and a photomultiplier tube (PMT) detector for fluorescence. Reagent addition is accomplished by an optional external injector module, which controls two independent injector syringes. Each syringe is connected to a separate injector tip. Emitted luminescence is captured using either the top or bottom probes and measured using the Synergy HT’s low noise PMT operated in photon integration mode. Here we describe the use of a Synergy™ HT Multi-Detection Microplate Reader (BioTek Instruments) to perform dual-luciferase measurements with purified recombinant enzymes.