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  2. LUCS (Light-Up Cell System), a universal high throughput assay for homeostasis evaluation in live cells

LUCS (Light-Up Cell System), a universal high throughput assay for homeostasis evaluation in live cells

  • Sci Rep. 2017 Dec 22;7(1):18069. doi: 10.1038/s41598-017-18211-2.
Sylvain Derick 1 Camille Gironde 1 Pierre Perio 2 Karine Reybier 2 Françoise Nepveu 2 Alain Jauneau 3 Christophe Furger 4
Affiliations

Affiliations

  • 1 Anti Oxidant Power AOP/MH2F-LAAS/CNRS, 7 avenue du Colonel Roche, BP 54200, Toulouse Cedex, 31031, France.
  • 2 Université de Toulouse, PHARMA-DEV, UPS-IRD, 118, route de Narbonne, Toulouse Cedex, 31062, France.
  • 3 CNRS, Institut Fédératif de Recherche 3450, Plateforme Imagerie, Pôle de Biotechnologie Végétale, Castanet-Tolosan, 31326, France.
  • 4 Anti Oxidant Power AOP/MH2F-LAAS/CNRS, 7 avenue du Colonel Roche, BP 54200, Toulouse Cedex, 31031, France. cfurger@laas.fr.
Abstract

Observations of fluorescent cyanine dye behavior under illumination at 500 nm lead to a novel concept in Cell Biology allowing the development of a new live cell assay called LUCS, for Light-Up Cell System, measuring homeostasis in live cells. Optimization of the LUCS process resulted in a standardized, straightforward and high throughput assay with applications in toxicity assessment. The mechanisms of the LUCS process were investigated. Electron Paramagnetic Resonance experiments showed that the singlet oxygen and hydroxyl radical are involved downstream of the LIGHT effect, presumably leading to deleterious oxidative stress that massively opens access of the dye to its intracellular target. Reversible modulation of LUCS by both verapamil and proton availability indicated that plasma membrane proton/cation antiporters, possibly of the MATE drug efflux transport family, are involved. A mechanistic model is presented. Our data show that intracellular oxidation can be controlled by tuning LIGHT energy, opening applications in regulatory purposes, anti-oxidant research, chemotherapy efficacy and dynamic phototherapy strategies.

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