Share Email Print
cover

Proceedings Paper

Two photon fluorescence imaging of lipid membrane domains and potentials using advanced fluorescent probes
Author(s): Vasyl Kilin; Zeinab Darwich; Ludovic Richert; Pascal Didier; Andrey Klymchenko; Yves Mély
Format Member Price Non-Member Price
PDF $14.40 $18.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

Biomembranes are ordered and dynamic nanoscale structures critical for cell functions. The biological functions of the membranes strongly depend on their physicochemical properties, such as electrostatics, phase state, viscosity, polarity and hydration. These properties are essential for the membrane structure and the proper folding and function of membrane proteins. To monitor these properties, fluorescence techniques and notably, two-photon microscopy appear highly suited due to their exquisite sensitivity and their capability to operate in complex biological systems, such as living cells and tissues. In this context, we have developed multiparametric environment-sensitive fluorescent probes tailored for precise location in the membrane bilayer. We notably developed probes of the 3-hydroxychromone family, characterized by an excited state intramolecular proton transfer reaction, which generates two tautomeric emissive species with well-separated emission bands. As a consequence, the response of these probes to changes in their environment could be monitored through changes in the ratios of the two bands, as well as through changes in the fluorescence lifetimes. Using two-photon ratiometric imaging and FLIM, these probes were used to monitor the surface membrane potential, and were applied to detect apoptotic cells and image membrane domains.

Paper Details

Date Published: 22 February 2013
PDF: 13 pages
Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85880S (22 February 2013); doi: 10.1117/12.2001492
Show Author Affiliations
Vasyl Kilin, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)
Zeinab Darwich, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)
Ludovic Richert, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)
Pascal Didier, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)
Andrey Klymchenko, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)
Yves Mély, Biophotonics and Pharmacology Lab., CNRS, Faculté de Pharmacie (France)


Published in SPIE Proceedings Vol. 8588:
Multiphoton Microscopy in the Biomedical Sciences XIII
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

© SPIE. Terms of Use
Back to Top