Prof. Véronique KRUYS, IBMM ULB
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.
Laure Twyffels, M.Sc. Bioengineering
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.
- Wide-field fluorescence microscope Zeiss Axio Observer Z1
• Inverted microscope with motorized stage
• Two light sources : LEDs or Xenon lamp DG4; numerous filtersets available
o 10x/0.3 EC Plan Neofluar Dry DIC ∞/- WD=5.2 mm
o 20x/0.5 EC Plan Neofluar Dry DIC ∞/0,17 mm WD=2.0 mm
o 20x/0.4 LD Plan Neofluar Dry DIC Corr ∞/0-1.5 mm WD=7.9 mm @0.75 mm
o 40x/1.3 EC Plan-Neofluar Oil Ph3 ∞/0.17 mm WD=0.21 mm
o 63x/1.4 Plan Apochromat Oil DIC ∞/0.17 mm WD=0.19 mm
o 100x/1.46 Alpha Plan-apochromat Oil DIC ∞/0.17 mm WD=0.11 mm
• Two cameras: high speed (Axiocam Hsm) or high resolution (Coolsnap HQ)
• Temperature-controlled incubation chamber with perfusion system
• Axiovision software with additional modules, including: Mark&Find, Time-lapse, Fast acquisition, Interactive Measurement, Z-stack and Deconvolution
- Laser-scanning confocal microscope Leica TCS SP2
• Inverted microscope
• 6 laser lines: 458, 476, 488, 514, 543, and 633 nm
o 10x/0.30 HC PL FL Dry ∞/- WD=11 mm
o 40x/0.55 N PLAN Dry PH2 ∞/Corr 0-2 mm WD=1.9-3.3
o 63x/1.40-0.60 HCX PL APO Oil ∞/0.17 mm WD=0,14 mm
o 100x/1.4 HCX PL APO Oil PH3 CS ∞/0.17 mm WD=0.09 mm
• Leica software for acquisition and basic processing
- Laser-scanning confocal microscope Zeiss LSM 710 NLO (multiphoton-ready):
• Inverted microscope with motorized stage
• 7 laser lines: 405 nm, 458 nm, 488 nm, 514 nm, 543 nm, 594 nm, and 633 nm
o 10x/0,3 EC Plan-Neofluar Dry DIC ∞/- WD=5,2 mm
o 20x/0,8 Plan-Apochromat Dry DIC ∞/0,17 mm WD=0,55 mm
o 40x/1,2 C-Apochromat Water DIC Corr ∞/0,14-0,19 mm WD=0,28 mm
o 63x/1,4 Plan Apochromat Oil DIC ∞/0,17 mm WD=0,19 mm
• HXP 120C metal halide lamp with filtersets for observation of common fluorochromes (DAPI (49), Cy3 (43), GFP (38), …)
• For an optimal control of the temperature (live imaging): Incubator XL multi S1 around the microscope and Heating insert PS1 inside to dock culture chambers into the stage. Possibility to control gas composition inside the incubator. ONIX Perfusion Microfluidic System from CellAsic.
• QUASAR 34-channels detector: 32-element array PMT detector and two flanking single PMT detectors, enabling acquisition of the entire visible spectrum as a λ-stack in one scan
• ZEN software with additional modules: Image VisArt plus (3D and 4D animation), ROI-HDR (high dynamic range imaging) and FRAP
1. A masked PY-NLS in Drosophila TIS11 and its mammalian homolog tristetraprolin. L. Twyffels, C. Wauquier, R. Soin, C. Decaestecker, C. Gueydan, and V. Kruys. PLoS ONE (in press)
2. dTIS11-dependent deadenylation is the key step in AU-Rich Element-mediated mRNA decay in Drosophila cells. C. Vindry, A. Lauwers, R. Soin, C. Wauquier, V. Kruys and C. Gueydan J. Biol. Chem. (Aug. 2012)
3. The HTLV-1 Tax protein inhibits formation of stress granules by interacting with histone deacetylase 6 (HDAC6). S. Legros, M. Boxus, JS. Gatot, C. Van Lint, V. Kruys, R. Kettman, JC. Twizere, and F. Dequiedt. Oncogene (2011) 30, 4050-4062
4. Pendrin : the thyrocyte apical membrane iodide transporter ? L. Twyffels, C. Massart, P.E. Golstein, E. Raspe, J. Van Sande, J.E. Dumont, R. Beauwens, R, V. Kruys. Cell. Physiol. Biochem. (2011) 28, 491-496.
5. SR proteins: more than splicing factors. L. Twyffels, C. Gueydan, and V. Kruys. FEBS Journal (2011) 278, 3246-3255.
6. 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. FEBS J. (2010) 277, 2496-2514.
7. FEBS J. 2011 Jul 27. doi: 10.1111/j.1742-4658.2011.08274.x.
8. The HTLV-1 Tax protein inhibits formation of stress granules by interacting with histone deacetylase 6 (HDAC6). S. Legros, M. Boxus, JC. Twizere, JS. Gatot, C. Van Lint, V. Kruys, R. Kettman, F. Dequiedt. Oncogene (2011) doi:10.1038.
9. 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.
10. 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.
11. 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.
12. 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.
13. 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
Description and applications
- 3D and time-lapse imaging and analysis of molecule subcellular distribution by direct fluorescence in living cells or by direct and indirect fluorescence in fixed cells and tissues.
- Co-localisation of fluorescent signals.
- Imaging and analysis of molecular interactions in living or fixed cells by FRET.
- Imaging and analysis of molecular diffusion by FRAP.
- Deconvolution of fluorescent signals and 3D-reconstruction.
See the poster: POSTER_CMMI_-_Fluorescence_Microscopy.pdf
See the "Molecular Biology of the Gene" website: http://www.ulb.ac.be/ibmm/homeuk_4.html