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C-ECi: A cubic-ECi combined clearing method for 3D follicular content analysis in The fish ovary†

Abstract : Deciphering mechanisms of oocyte development in female fishes still remains challenging and a comprehensive overview of this process at the level of the organ is still needed. The recent development of optical tissue clearing methods has tremendously boosted the 3D imaging of large size biological samples that are naturally opaque. However, no attempt of clearing on fish ovary that accumulates extremely high concentration of lipids within oocytes has been reported to date. To face with this ovarian-specific challenge, we combined two existing clearing methods, the non-toxic solvent-based ECi method for efficient clearing and the CUBIC method to enhance lipid removal and reduce non-specific staining. The methyl green fluorescent dye was used to stain nuclei and delineate the follicular structures that include oocytes. Using this procedure (named C-ECi), ovaries of both medaka and trout could be imaged in 3D and follicles analyzed. To our knowledge this is the first procedure elaborated for clearing and imaging fish ovary in 3D. The C-ECi method thus provides an interesting tool for getting precise quantitative data on follicular content in fish ovary and promises to be useful for further developmental and morphological studies.
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https://hal.inrae.fr/hal-02935099
Déposant : Maryse Corvaisier <>
Soumis le : jeudi 10 septembre 2020 - 09:00:24
Dernière modification le : mercredi 16 septembre 2020 - 09:20:04

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Manon Lesage, Manon Thomas, Jérôme Bugeon, Adèle Branthonne, Stéphanie Gay, et al.. C-ECi: A cubic-ECi combined clearing method for 3D follicular content analysis in The fish ovary†. Biology of Reproduction, Society for the Study of Reproduction, 2020, ⟨10.1093/biolre/ioaa142⟩. ⟨hal-02935099⟩

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