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  Director : Marie-Christine PRÉVOST (mprevost@pasteur.fr)


  abstract

 

The electron microscopy platform serves three main areas of activity in electron microscopy. The first concerns the development and application of analytical ultra-structural tools for cellular biology, virology and microbiology in studies requiring both standard electron microscopy procedures as well as specialised techniques, such as immuno-gold labelling and cryopreservation and fixation methods. The second activity pertains to quality control of different biological samples (cell lines, primary cells, viral, bacterial and fungal isolates), while the third area centres on teaching and training activities.



  report

cale

The electron microscopy platform is a laboratory whose mission is to serve the Institut Pasteur community by providing technical and support resources for studies at the ultra-structural level. The laboratory is equipped with several transmission electron microscopes (Philips CM10 and CM12, Jeol JEM 1010 and 1200 EX) and, since November 2002, with a JSM-6700F scanning electron microscope with a cold-emission cathode "cold-FEG". This instrument, which enables a resolution of 2-3 nm at 1 KV for biological specimens, should become fully operational in the first semester of 2003. The laboratory also offers a wide spectrum of materials and equipment for the preparation and analysis of biological samples, and training for researchers and technical staff in the autonomous use of the laboratory's resources.

Furthermore, in accordance with the guidelines set by Institut Pasteur's strategic council (Conseil de Stratégie d'Orientation) and the coordinator of technical platforms, the laboratory will play an active role in the development of innovating methodologies, such as cryo-preparation techniques for cell biology applications.

Work within the platform is conducted on a collaborative contract basis, under which the objectives, possibilities, technical orientation and risks, as well as confidentiality and recognition-related issues, are defined. The recently created "supervising committee" validates such collaborations, which might involve several different laboratories and/or research groups.

Within this framework, several collaborative projects have been initiated with various research units of the institute. Seven of the 15 projects initiated in 2001 have been completed and 26 projects were initiated in 2002. Examples of on-going collaborations are listed below.

Virology

Studies on HIV: Entry of the virus into primary cells : lymphocytes, macrophages and dendritic cells. Interaction of HIV with nucleolin at the cell surface and interference with the pseudopeptide HB19.

Studies on VHC: Immunocaracterisation of viral pseudoparticles (VLPs) by negative staining and visualisation of their cellular localisation.

Studies on HTLV-1: Visualisation of chimeric HTLV/MuLV particles in an experimental model of murine viral infection. Study of the GAG protein during budding of the HTLV-1 retrovirus.

Molecular Medicine Studies on HIV: Role of the mutations of the DNA flap in the function of the pre integration complex.

Microbial PathogenesisSubcellular localisation of various antigenic proteins involved in the virulence of Mycobacterium tuberculosis, Mycobacterium bovis, BCG and Mycobacterium segmatis.

Structure and Dynamic of GenomesMorphological analysis of Aspergillus fumigatus conidia of cell wall-mutants and their interaction with the alveolar macrophage.

Immunology: Effect of mosquito salivary antigens on the degranulation of mastocytes and their role in the induction of infections by Plasmodium falciparum.

ParasitologyStudies of the nuclear organisation in the human pathogen Plasmodium falciparum : Immunolocalisation of telomerase.

Ecosystems and Epidemiology of infectious diseases.: Interaction between Bordetella pertussis and human tracheal cells : Ultrastructural analysis of invasive potential and intracellular survival of Bordetella pertussis, Bordetella parapertusis and Bordetella bronchiseptisca.

Expertise and external collaborations

We routinely perform analyses of different biological samples for the presence of virus, bacteria and fungi in various cell lines or primary cell cultures (investigations requested by campus scientists). An ongoing research contract with Texcell provides services for external companies. The many research collaborations that have been developed with scientists outside of the institute include investigations of the role of mitochondria in apoptosis and identification and cytolocalisation of surface antigens in Mycoplasma.

Teaching

Microscopy techniques are continuously taught to scientists and technicians involved in electron microscopy investigations. This will serve as a basis for setting up a new Electron Microscopy Course (Formation continue Pasteur, 27, 28 January 2003).

Intramural Collaborations

Viruses and immunity (N. Sol, O. Schwartz)

Slow Viruses (F. Tangy, M. Brahic)

Molecular Genetics (G. Vignon, A. Pugsley)

Virology and Cellular Immunology (S. Nisole, A. Hovanessian)

Molecular Genetics of Respiratory Tracts Viruses (A. Martin, S. Van der Werf)

Bacterial Molecular Genetics (N. Honoré, S. Cole)

Mycobacterial Genetics (O. Neyrolles, B. Gicquel)

Pathogenesis of Mucosal Bacteria (H. De Reuse, A. Labigne)

Retroviruses and Gene Transfer (N. Desmaris, J.M. Heard)

Aspergillus (J.P. Latgé)

Immunoallergology (C. Demeure, S. Mecheri)

Lympho-Epithelial interaction (S. Kerneis, M. Marchetti, E. Pringault)

Molecular Virology and Vectorology (P. Charneau)

Photo : Mycobacterium Tuberculosis Strains "H37Rv", grown in 7H9 medium, analysed by scanning electron microscopy on Jeol JSM 6700 F

Keywords: Electron Microscopy, Ultrastructur, Immunogold, Cryo sections, Cryo fixation



  publications

puce Publications of the unit on Pasteur's references database


  personnel

  Office staff Researchers Scientific trainees Other personnel
  Danièle LHOUMEAU, dlhoum@pasteur.fr   Michèle TORRES Marie-Christine PRÉVOST, mprevost@pasteur.fr

Isabelle BONNE, ibonne@pasteur.fr

Christian BRONNERT, cbronner@pasteur.fr

Brigitte CHAVINIER-JOVÉ, bchavijo@pasteur.fr

Stéphanie GIROUX, sgiroux@pasteur.fr

Christine SCHMITT, cschmitt@pasteur.fr

Nadège CAYET


Activity Reports 2002 - Institut Pasteur
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