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    4. PICT: APPLICATIONS

    APPLICATIONS
     

    We are happy to share expertise in scientific projects addressing defined questions which can benefit from: (i) electron microscopy (scanning and transmission), including ultrastructural cytochemistry; (ii) high-resolution and high-throughput confocal microscopy, for live-cell imaging and immunolabelling; (iii) multiphoton microscopy to image tissues and organs for several hours without damage; and (iv) advanced applications by confocal microscopy, including dynamics of molecular movement by Fluorescence Recovery After Photobleaching (FRAP), protein dimerization by Bimolecular Fluorescence Complementation (BiFC), in situ detection of free radicals (ROS) at distinct subcellular compartments, and protein:protein interactions by FRET, etc. In addition, we propose advanced images analyses such as deconvolution, 3-D reconstruction, morphometry, objective co-localization, etc.

     

    EXPERTISES

     

    - Basic training and supervision of routine imaging

    		  

    - Sophisticated methods for preparation of biological samples (cells, tissues and organs)

    - Advanced training destined for independent, registered platform users

    		  

    - Vital cell and subcellular imaging with particle tracking

    - Fluorescence imaging on fixed cells and tissues.
     

    Salivary glands explants (unpublished data).

     

     

     

     

     




    Acquired with Stereomiscroscope Discovery V1

    		  

    - Extended tissue and organ imaging without damage by multiphoton microscopy

    - High-resolution confocal imaging on fixed cells, tissues and organs.
     

    Triple (immuno)labelling of fibroblasts.

    Acquired with LSM510

     

    		  

    - Fluorescence imaging on living cells: cell migration. 
     

    Migration of epithelial cells (Platek et al, 2004).


    Acquired with Axiovision

    - High-throughput confocal microscopy.

    Triple (immune)labelling. 

     
    Acquired with COSD

     

    		  

    - High-resolution confocal imaging on living cells. 

    Vital imaging of stable submicrometric membrane lipid domains (D’auria et al, 2013).

     
    Acquired with LSM510

     

    - Advanced applications by confocal microscopy:

    • Dynamics of molecular movement (Fluorescence Recovery After photobleaching: FRAP).


    Acquired with LSM510

    • Protein:protein interaction (Forster resonance energy transfer: FRET). TCR/CD8 interaction (Demotte et al, 2010).


    Acquired with LSM510

    • Protein dimerization by fluorescence complementation: BiFC.


    Acquired with LSM510

    • In situ detection of free radicals (ROS). Differential ROS localization (Denamur S*, Tyteca D* et al, 2011).


    Acquired with LSM510

    • In situ analysis of membrane:drug interaction.
    • Dynamics of lipid and protein membrane domains.
    • Ratiometric microscopy.
    		  

    - Electron microscopy:

    • SEM (scanning electron microscopy). Featureless membrane of red blood cells (Carquin et al, in preparation).

     
    Acquired with LSM510

    • TEM (transmission electron microscopy)
    • STEM (combined TEM/SEM)
    • Ultrastructural cytochemistry
    • Ultrastructural colloidal gold labelling High-resolution protein distribution: immunogold surface labelling: ASGP receptors on hepatocytes (Van Der Smissen et al, 1996).


    Acquired with TEM-CM12

    • Ultrastructural analysis of particles (large protein complexes, viruses, liposomes)

     


     

    		  

    - Advanced off-line image analysis

    • Size distribution Liposomes (Fa et al, 2007).


    Acquired with Axiovision

     

    • Colocalization analysis using line intensity profiles Veiga-Da-Cunha et al, 2010.


    Acquired with LSM510

     

    • 3D-deconvolution and reconstruction


    Acquired with COSD

     

     

    PICT

    Platform for Imaging Cells and Tissues