Bioluminescence technology is developing as an important tool in biological research. Borrowed from species of jellyfish and firefly, luciferase genes are being used to create bioluminescence in cells not normally bioluminescent. This technology can be used in applications such as gene reporters, microarray analyses, and real-time monitoring of protein interactions. Two luciferases with a depth of characterization and widely used in cellular and molecular biology are Firefly luciferase (Fluc), isolated from Photinus pyralis, and Renilla luciferase (Rluc), isolated from Renilla reniformis.;The emission wavelengths of Fluc and Rluc differ such that they are distinguishable using appropriate optical filters. Technology that is capable of monitoring light emission of two luciferases simultaneously has the potential to benefit a wide range of biomedical research areas. In this study a novel apparatus has been tested to determine the capability and quality of monitoring light emission from Fluc and Rluc simultaneously. Assays were developed to serve as examples of potential uses for the dual-luciferase monitoring apparatus. Using cell lines modified with luciferase genes, it was possible to show that the apparatus could successfully distinguish between light emission from Fluc and Rluc, monitor luciferase activity in a 3-D mimic of an in vivo environment, and quantify changes in a biosensor-type cell line expressing an additional luciferase as an internal control.
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