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

Sergi Padilla-Parra 1 Nicolas Audugé 1 Maïté Coppey-Moisan 1, * Marc Tramier 1, 2, *
* Auteur correspondant
1 IJM - Institut Jacques Monod
2 IGDR - Institut de Génétique et Développement de Rennes
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.
Keywords : FCS FCCS FLCS quantitative fluorescence microscopy TCSPC
Type de document :
Article dans une revue
Microscopy Research and Technique, Wiley, 2011, 74 (8), pp.788-93. 〈10.1002/jemt.21015〉
Liste complète des métadonnées

Littérature citée [28 références]  Voir  Masquer  Télécharger
http://www.hal.inserm.fr/inserm-00604664
Contributeur : Hervé De Villemeur <>
Soumis le : jeudi 30 juin 2011 - 12:01:10
Dernière modification le : mercredi 16 mai 2018 - 11:22:33
Document(s) archivé(s) le : lundi 5 décembre 2016 - 00:54:33

Fichier

Padilla-Parra_MRT-1.pdf
Fichiers produits par l'(les) auteur(s)

Identifiants

Collections

IJM | IGDR | BIOSIT | UR1-UFR-SVE | USPC | UNIV-RENNES

Citation

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, Wiley, 2011, 74 (8), pp.788-93. 〈10.1002/jemt.21015〉. 〈inserm-00604664〉

Exporter

BibTeX TEI DC DCterms EndNote

Partager

Métriques

Consultations de la notice

533

Téléchargements de fichiers

586

Contact

support.ccsd.cnrs.fr
hal.support@ccsd.cnrs.fr
Logo CCSD