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Pr Véronique Kruys, responsable (ULB, IBMM)
vkruys@ulb.ac.be

Véronique Kruys was born in 1963 and studied at the Université Libre de Bruxelles (ULB), Belgium, from which she graduated with a Lic. Sci. Degree in 1985. She obtained her Ph.D. in 1989 from the same University while she was appointed Assistant. During her thesis, she identified a major mechanism controlling cytokine gene expression at the post-transcriptional level which was reported in papers published in PNAS and Science. She went on post-doc for three years (1990-1002) in the Laboratory of Bruce Beutler, at the Howard Hughes Medical Institute, University of Texas at Dallas. On her return at the Free University of Brussels, she was appointed Assistant Professor at the Department of Molecular Biology of the Faculty of Sciences. Her scientific interest remained focused on biological processes controlling messenger RNA metabolism. She published around 40 publications and several reviews. She has been teaching biochemistry and molecular biology of genes for several years. In the recent years, her research interests focused on the characterization of protein and RNA traffic which led her to develop her expertise in fluorescence microscopy.

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Laure Twyffels, M.Sc. Bioengineering
ltwyffel@ulb.ac.be

Born in 1984, Laure Twyffels obtained her Master degree in bioengineering from the Université Libre de Bruxelles (ULB) in 2007. She then started a PhD at the Institute of Molecular Biology and Medecine of the ULB as a Research Fellow of the Belgian National Fund for Scientific Research. Her PhD thesis is focused on the nucleocytoplasmic transport of two RNA-binding proteins involved in ARE-mediated post-transcriptional regulation of gene expression. Through her research, she has gained expertise in molecular biology, fluorescence microscopy, live imaging and immunofluorescence techniques. Since October 2011, she works for the Fluorescence Microscopy Unit within the CMMI, where she provides training, technical and scientific support for the users of the facility.

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Liste des axes > Microscopie à fluorescence

Microscopie à fluorescence

 

Équipements

  1. Microscope à fluorescence Zeiss Axio Observer Z1 inversé, équipé d’une plateforme motorisée, d’une chambre d’incubation thermostatisée avec système de perfusion, d’une caméra digitale à haute vitesse (Axiocam Hsm) et d’une caméra digitale à haute résolution (coolsnap HQ). Double source lumineuse Colibri/Lambda DG4.
  2. Microscope confocal inversé Leica TCS SP2 (λ d’excitation : 458, 476, 488, 514, 543, 633 nm)
  3. Microscope confocal à balayage laser Zeiss LSM 710 NLO (multiphoton ready):
    • Microscope inversé à plateforme motorisée
    • 7 lignes laser : 405 nm, 458 nm, 488 nm, 514 nm, 543 nm, 594 nm et 633 nm
    • Objectifs :
    o 10x/0,30 EC Plan-Neofluar WD=5,2 mm DIC ∞/- D=0,17 mm
    o 20x/0.80 Plan-Apochromat WD=0,55 mm DIC ∞/- D=0,17 mm
    o 40x/1.20 C-Apochromat Water DIC Corr D=0,14-0,19 mm
    o 63x/1.40 Plan-Apochromat Oil DIC D=0,17 mm
    • Lampe aux halogénures métalliques HXP 120C avec filtres permettant la détection des fluorochromes communément utilisés (DAPI (49), Cy3 (43) et GFP (38), …)
    • Pour un contrôle optimal de la température des échantillons vivants : incubateur XL multi S1 autour du microscope et adaptateur chauffant P S1  pour chambres de culture. Possibilité de contrôle des gaz à l’intérieur de l’incubateur.
    • Détecteur QUASAR 34-canaux : 2 photomultiplicateurs simples encadrent une rangée de 32 photomultiplicateurs, ce qui permet l’acquisition de tout le spectre (λ-stack) en un seul balayage du champ.
    • Logiciel ZEN avec modules additionnels
    o Image VisArt plus (animation 3D et 4D)
    o ROI HDR (imagerie à grande gamme dynamique)
    o FRAP
    o FRET

Références bibliographiques

  1. Pendrin: the Thyrocyte Apical Membrane Iodide Transporter? L. Twyffels, C. Massart, P. E. Golstein, E. Raspe, J. Van Sande, J.E. dumont, R. Beauwens, V. Kruys. Cell Physiol Biochem 2011;28:491-496, Oct. 17,2011
  2. Shuttling SR proteins: more than splicing factors. Twyffels L, Gueydan C, Kruys V.

    FEBS J. 2011 Jul 27. doi: 10.1111/j.1742-4658.2011.08274.x.

  3. The HTLV-1 Tax protein inhibits formation of stress granules by interactin with histone deacetylase 6. Legros S, Boxus M, Gatot JS, Van Lint C, Kruys V. Kettmann R, Twizere JC, Dequiedt F. Oncogene. 2011 May 2. 

  4. The splicing factor ASF/SF2 is associated to TIAR/TIA-1-containing ribonucleoproteic complexes and contributes to post-transcriptional repression of gene expression. N. Delestienne, C. Wauquier, R. Soin, J.-F. Dierick, C. Gueydan and V. Kruys (last 2 authors equally contributed). FEBS J. (2010) 277, 2496-2514.
  5. Impaired embryonic development in mice overexpressing the RNA-binding protein TIAR. Y. Kharraz, P.-A. Salmand, A. Camus, J. Auriol, C. Gueydan, V. Kruys, D. Morello. PLoS ONE (2010) 5(6) e11352.
  6. Identification of the sequence determinants mediating the nucleo-cytoplasmic shuttling of TIAR/TIA-1 RNA-binding proteins. T. Zhang, N. Delestienne, G. Huez, V. Kruys, and C. Gueydan. J. Cell Sci. (2005) 118, 5453-5463.
  7. The Vacuole System Is a Significant Intracellular Pathway for Longitudinal Solute Transport in Basidiomycete Fungi. P.R. Darrah, M. Tlalka, A. Ashford, S.C. Watkinson, and M.D. Fricker. Eukaryotic Cell (2006) 5, 1111-1125.
  8. Intracellular NAD levels regulate TNF- protein synthesis in a sirtiun-dependent manner. F. Van Gool, M. Galli, C. Gueydan, V. Kruys, P.-P. Prévot, A. Bedalov, R. Mostoslavsky, F. Alt, T. De Smedt and O. Leo. Nature Medicine (2009) 15, 206-210.
  9. Post-transcriptional regulation of genes encoding anti-microbial peptides in drosophila. A. Lauwers, L. Twyffels, R. Soin, C. Wauquier, V. Kruys and C. Gueydan. J. Biol. Chem. (2009) 284, 8973-8983

Descriptif et applications

  1. Analyse de la distribution subcellulaire  de molécules en 3 dimensions par fluorescence directe dans des cellules vivantes (temps réel) ou par fluorescence directe ou indirecte sur échantillons de cellules ou de tissus fixés.
  2. Analyse de co-localisation de signaux fluorescents
  3. Analyse d’interactions moléculaires en temps réel par FRET
  4. Analyse de processus de diffusion moléculaire par FRAP
  5. Déconvolution de signaux fluorescents et reconstitution en 3 dimensions.

Voir le poster: POSTER_CMMI_-_Fluorescence_Microscopy.pdf

Voir le site "Biologie moléculaire du gène" : http://www.ulb.ac.be/ibmm/homeuk_4.html

 

 
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