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Deconvolution of luminescence cross-talk in high-throughput gene expression profiling.

08:00 EDT 16th May 2019 | BioPortfolio

Summary of "Deconvolution of luminescence cross-talk in high-throughput gene expression profiling."

Luciferase reporters have become standard genetic tools to monitor gene expression in real time and in high-throughput using microplate readers. Compared to reporter gene assays based on fluorescence proteins, luciferase reporters have a superior signal-to-noise ratio, since they do not suffer from the high auto-fluorescence background of the bacterial cell. However, at the same time luciferase reporters have the drawback of constant light emission, which leads to undesired cross-talk between neighbouring wells on a microplate. To overcome this limitation, we developed a computational method to correct for luminescence bleed-through and to estimate the "true" luminescence activity for each well of a microplate. As the sole input our algorithm uses the signals measured from a calibration plate, in which the light emitted from a single luminescent well serves as an estimate for the "light-spread function". We show that this light-spread function can be used to deconvolve any other measurement obtained under the same technical conditions. Our analysis demonstrates that the correction preserves low-level signals close to the background and shows that it is universally applicable to different kinds of microplate readers and plate types.

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This article was published in the following journal.

Name: ACS synthetic biology
ISSN: 2161-5063
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