New tool to monitor membrane potential by FRET Voltage Sensitive Dye (FRET-VSD) using Spectral and Fluorescence Lifetime Imaging Microscopy (FLIM). Interest in cell engineering
Issue title: Selected Papers from the First Seminar on “Medical Engineering and Therapy”, 27–28 April 2004, Nancy, France
Affiliations: Laboratoire de Mécanique et Ingénierie Cellulaire et Tissulaire, UMR CNRS 7563 LEMTA et IFR 111 CNRS – UHP-INPL-CHU, 54505 Vandoeuvre lès Nancy, France
Note: [] Corresponding author: Dominique Dumas, Plateau Technique d'Imagerie et de Biophysique Cellulaire et Tissulaire de Lorraine, Laboratoire de Mécanique et Ingénierie Cellulaire et Tissulaire, UMR CNRS 7563 et IFR 111, Faculté de Médecine, BP 184, 54505 Vandoeuvre lès Nancy cédex, France. Tel.: +33 03 83 68 34 65; Fax: +33 03 83 68 34 59; E-mail: Dumas@medecine.uhp-nancy.fr.
Abstract: In this work, we investigated a voltage-sensitive fluorescent system to monitor membrane potential by spectral and lifetime fluorescence microscopy. A two-component FRET sensor has been designed that utilizes fluorescent phospholipids acceptor (DHPE-TRITC) bound on one side of the membrane and donor molecules (oxonol) which are sensitive to membrane potential. We used multiphoton excitation and FLIM to deliver contrast lifetimes of different line cancerous cells. These results provide new information concerning the differential response to depolarized cancerous cells from resting cells when compared to fibroblast normal cells. Given the sensitivity and the fast time response, this FRET system may be particularly useful for applications involving compression of tissues by mechanical forces.
Keywords: Potentiometric dyes, FLIM, spectral, oxonol, valinomycin, fluorescence resonance energy transfer
