Book of Abstracts - Ruhr-Universität Bochum

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OP-39 ISBOMC `10 5.7 – 9.7. 2010 Ruhr-Universität Bochum Subcellular Imaging of a Re(CO)3 Complex by Photothermal Infrared Spectromicroscopy (PTIR). Anne Vessières, a Clotilde Policar, b Marie-Aude Plamont, a Sylvain Clède, b Alexandre Dazzi c a ENSCP, CNRS-UMR 7223, 11 rue P. et M. Curie, F-75231 Paris Cedex 05, France, b Département Chimie de l’ENS, CNRS-UMR 7203, 24 rue Lhomond, F-75231 Paris Cedex 05, c Laboratoire de Chimie Physique, CNRS-UMR 8000, Université Paris-Sud 11, F-91405 Orsay Cedex, E-mail: a-vessieres@chimie-paristech.fr The most widely developed techniques for bio-imaging are those based on fluorescence spectroscopy, however vibrational techniques including infra-red (IR) are also valuable. However, in classical optical microscopy, sub-micrometric resolutions are not attainable in the mid-IR-range, as the diffraction criterion imposes resolution higher than �/2 (i.e. 2.5 µM at 2000 cm -1 ) which is not well suited for intracellular mapping. To reach sub-micrometric resolution, near-field techniques are mandatory. Photothermal induced resonance (PTIR) is a cutting edge technique using a set-up recently patented by Dazzi et al., 1 coupling atomic force microscopy (AFM) and a tunable infrared laser to make spatially resolved absorption measurements in the IR-range. It has been successfully used to map a single air-dried E. coli cell by irradiation in the amide I and II bands. 2 The next challenge is the identification and localization of exogeneous diluted compounds inside single cells. The Re(CO)3 unit grafted to a hydroxy-tamoxifen-like molecule has been selected for this study as it is stable in biological environments and displays intense absorption in the 1850 – 2100 cm -1 region where biological samples are transparent. Figure : left: AFM-set up. middle: PTIR mapping at 1925 cm -1 of a single cell incubated 1h with 10 µM of the Re(CO)3 complex (red = high concentration) right : spectromicroscopy at the nucleus We will present the results of the chemical imaging of this Re complex, inside a single cell, using PTIR. Cells are initially located on the surface of a ZnSe prism by using the AFM topology. Then the complex is localized thanks to its two characteristic �CO bands at 1925 and 2017 cm -1 . Cells showed an uneven distribution of the complex with one hot spot (red area on the picture above) and cold regions (blue zones). Interestingly, this location seems to correspond to cell nucleus located using irradiation at 1240 cm -1 (phosphate band) and 1650 cm -1 (amide I of proteins). In addition, a spectrum recorded inside the hot spot shows the two characteristic bands of the complex at 1925 and 2017 cm -1 , thus confirming its presence (right part of the figure). References 1. A. Dazzi, M. Reading, P. Rui, K. Kjoller, Patent 2008, WO/2008/143817 A. B. Lastname, C. D. 2. A. Dazzi, R. Prazeres, F. Glotin, J.-M. Ortega, Infrared Physics Techn. 2006, 49, 113. 55
OP-40 ISBOMC `10 5.7 – 9.7. 2010 Ruhr-Universität Bochum Dynamical Studies of Bioconjugated Luminescent Ruthenium Complexes in Lipid Vesicles Edward Rosenberg, a Ayesha Sharmin, a J. B. Alexander Ross a a Department of Chemistry and Biochemistry University of Montana, Missoula, MT 59812, USA Email: edward.rosenberg@mso.umt.edu A detailed analysis of the time-resolved anisotropy decay of the emission from luminescent molecules can provide useful information about molecular dynamics in a given media. To obtain such information, the excited-state lifetime must match the time scale of the process being examined and it is desirable that the time-zero emission anisotropy be in the range of 0.1 or greater. In our prior work, we designed a series of phosphorescent ruthenium complexes that have the longer lifetimes, higher quantum yields and the lower symmetry required for studying the dynamics of these probes in lipid vesicle bilayers. 1 Starting with the complexes [Ru(H)(trans-PPh3)2(dcbpy)CO][PF6] (1) and [Ru( dppene) (5-amino- phen)CO(TFA)][PF6] (2) (dcbpy = 4,4’-dicarboxy bipyridine, dppene = 1,2diphenylphosphino ethene, 5-amino phen = 5-amino phenanthroline, TFA = trifluoroacetic acid) we have used standard techniques to covalently conjugate these molecules to two and one phosphatidyl ethanolamine molecules, respectively. Complex 2 has also been conjugated to cholesterol via reaction with cholesterol chloroformate. Analyses of the anisotropy decay as a function of temperature from the conjugated probes in vesicles—made from both naturally occurring and synthetic lipids—reveal that the lipid conjugates are located within the lipid bilayer while the cholesterol conjugated complex is at the bilayer-water interface. The rotational correlation times for the complexes are responsive to the nature of the lipid vesicle used and analyses of this parameter allowed a detailed picture of the kinds of motions of the conjugated complex in the vesicle. The lipid conjugates with 1and 2 incorporated into vesicles via extrusion through a size-selective membrane, showed an unexpected blue-shift in the emission spectra and a corresponding short excited-state lifetime in the range of 10 ns; the lifetimes in solvents are in the range of 0.1-2 �s. This unusual phenomenon will be discussed in terms of the properties of the excited states in the lipid vesicle environment in contrast to in bulk solvents. For comparison with complexes 1and 2, related complexes have been conjugated to lipids and cholesterol via the phosphine ligands, and their photophysical properties in lipid vesicles will also be presented References 1. Ayesha Sharmin , Reuben C. Darlington , Kenneth I. Hardcastle, Mauro Ravera, Edward Rosenberg, J. B. Alexander Ross “Tuning Photophysical Properties with Ancillary Ligands in Ru(II) Mono- Diimine Complexes,” J. Organometal. Chem. 2009, 694, 988-1000. 56
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Sergey Abramkin Universität Wien,
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Tensim Dallagi ENSCP, France t.dall
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Prof. Dr. Toshikazu Hirao Osaka Uni
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Obiora Augustine Orakwue Hyqid Nige
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Book of Abstracts - Ruhr-Universität Bochum

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