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Healthy follicles were identified by an intact basal lamina, a visible oocyte with a germinal vesicle, and a distinct nucleolus, while atretic follicles were characterized by pyknotic nuclei and degenerating granulosa cells.23 Immunohistochemistry The immunohistochemical staining kit (Thermo Ultravision Detection System, Fremont, USA, TP-125-HL) was used in conjunction with the streptavidin-biotin-peroxidase method

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