Biological Engineering Commons^™

Bioluminescence Imaging For Assessment And Normalization In Transfected Cell Arrays, Angela K. Pannier, Eric A. Ariazi, Abigail D. Bellis, Zain Bengali, V. Craig Jordan, Lonnie D. Shea

Department of Biological Systems Engineering: Papers and Publications

Transfected cell arrays (TCAs) represent a high-throughput technique to correlate gene expression with functional cell responses. Despite advances in TCAs, improvements are needed for the widespread application of this technology. We have developed a TCA that combines a two-plasmid system and dual-bioluminescence imaging to quantitatively normalize for variability in transfection and increase sensitivity. The two-plasmids consist of: (i) normalization plasmid present within each spot, and (ii) functional plasmid that varies between spots, responsible for the functional endpoint of the array. Bioluminescence imaging of dual-luciferase reporters (renilla, firefly luciferase) provides sensitive and quantitative detection of cellular response, with minimal post-transfection processing. …