KEYWORDS: Electron microscopy, Nanoparticles, Luminescence, Microscopy, Platinum, Scanning electron microscopy, Absorption, Magnesium, Chemical species, Iron
Nowadays, many crucial biological questions involve the observation of biological samples at different scales. Thus, optical microscopy can be associated to magnetic nuclear imaging allowing access to data from the cellular to the organ level, or can be associated to electron microscopy to reach the sub cellular level. We will describe here the design, synthesis and characterization of new bimodal probes, which can be used as dye in two-photon excited microscopy (TPEM) and electron dense markers in scanning and transmission electron microscopy (EM). In a first part, we will describe new molecular dyes with small organic systems grafted on metal atoms (Pt, Au). Such systems show good twophoton induced fluorescence and two-photon images of HeLa cells will be presented. In a second part, we will present hybrid organic-inorganic fluorescent systems with diketopyrrolopyrole-based dye grafted on iron oxide-silica core shell nanoparticles by peptide bond. Such systems present high two-photon absorption cross sections and good fluorescence quantum yields. These nanoparticles are rapidly internalized in HeLa cells and high quality two-photon images were performed with low laser power. Then we will present our results on correlative light-electron microscopy were twophoton and electron microscopy (both scanning and transmission) images were obtained on the same biological sample.
We describe the molecular engineering of new highly efficient two-photon photolabile cages, based on dissymmetrical
donor-acceptor and symmetrical donor-donor systems. Complete characterizations of these new cages, based on a
biphenyl, fluorenyl or a 2,5-diphenylthienyl central core will be presented. In vitro photochemistry will be described
(uncaging quantum yield, two-photon uncaging action cross-section,...) and we will then focus on biological results
obtained in cell culture and on acute brain slices (with glutamate and fluorescence uncaging).
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