Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. - Archive ouverte HAL Access content directly
Journal Articles Microscopy Research and Technique Year : 2011

Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell.

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Abstract

Dual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with crosstalk were overcome with an approach based on fluorescence lifetime correlation spectroscopy (FLCS). We show that FLCS applied to dual-color EGFP and mCherry cross-correlation allows the determination of protein-protein interactions in living cells without the need of spectral bleed through calibration. The methodology was validated by using EGFP-mCherry tandem in comparison with coexpressed EGFP and mCherry in live cell. The dual-color FLCS experimental procedure where the different laser intensities do not have to be controlled during experiment is really very helpful to study quantitatively protein interactions in live sample.
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Dates and versions

inserm-00604664 , version 1 (30-06-2011)

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Sergi Padilla-Parra, Nicolas Audugé, Maïté Coppey-Moisan, Marc Tramier. Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell.. Microscopy Research and Technique, 2011, 74 (8), pp.788-93. ⟨10.1002/jemt.21015⟩. ⟨inserm-00604664⟩
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