Tissue clearing and immunostaining

               Natural opacity of tissues prevents 3D imaging of big samples. Emergence of clearing methods dedicated to various tissues have been developed with the aim to render tissues transparent for high resolution imaging of thick tissues. We tested and implemented several clearing approaches on zebrafish organisms (from larvae to juvenile and adult) allowing 3D imaging of whole-organism at (sub)cellular resolution. We routinely use simple immersion methods, hyperhydration methods and solvent-based method combined with 2-photon or light-sheet imaging (Institut de la vision - imaging facility). These state-of-the-art sample preparation and imaging techniques allow a unique view of infectious processes at different stages of zebrafish development, morphological characterization of deep tissues and detailed quantification at the whole organ level. We further extend our expertise to various vertebrate tissues: medaka (ovaries), trout (spleen, ovaries), mouse (lung, nasal cavity, brain, …), pig (skin, lymphoid ganglia).

Collaboration with Betrand Bryche and Nicolas Meunier, UVSQ / INRA Jouy-en-Josas.

Collaboration V. Thermes, LPGP, INRA Rennes

Collaboration V. Beringue & JL. Vilotte, INRA Jouy en Josas

Collaboration P. Hernandez & JP. Levraud, Institut Pasteur PARIS

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