Low-cost system for real-time monitoring of luciferase gene expression.
In some mammalian cells transfected with luciferase reporter genes, the luciferase/luciferin reaction in a cell monolayer produces a very small light flux. While the low light levels are often measurable with single-photon counting cameras, these devices are expensive and may require long averaging times to acquire an image. We describe an approach for real-time monitoring of light produced by luciferase gene expression in intact, cultured cells using readily available and relatively inexpensive components. The system uses a single-photon counting photomultiplier tube with built-in high voltage supply and photon counting circuitry to rapidly measure average light output from growing cells in a 35 mm culture dish. The fast, accurate and highly sensitive response of the system makes it useful for studying the dynamics of gene expression over time periods ranging from minutes to days.
Gailey
P C
PC
Energy Division, Oak Ridge National Laboratory, TN 37831, USA. pg7@stc06.ctd.ornl.gov
Miller
E J
EJ
Griffin
G D
GD
D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D002478:Cells, Cultured; D004958:Estradiol / Q000494:pharmacology; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D008027:Light; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D010783:Photometry / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D017785:Photons; D013629:Tamoxifen / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D018931:Antineoplastic Agents, Hormonal; D013629:Tamoxifen; D004958:Estradiol; D008156:Luciferases