Tissue was collected from sexually mature mice (>8 weeks old).
Mlh3−/− mice (The Jackson Laboratory; #018845) were bred at the Washington University in Saint Louis and processed as previously described (Jung et al., 2019 (link)). Mice of the Crispy line were bred by the Ahituv laboratory at the University of California, San Francisco, and fixed testes were received in 70% ethanol. Wild-type C57BL/6J (The Jackson Laboratory; #000664) were bred in-house at the Small Laboratory Animal Unit (ONPRC/OHSU). All animal experiments were performed in compliance with the regulations of the respective host institutions.
To capture subtle image differences resulting from experimental variables, the samples used to train the neural network were processed under different conditions: (i) fixation method—perfusion or immersion; (ii) fixative type—4% paraformaldehyde (PFA) or modified Davidson’s fixative and (iii) assay—IF with or without Tyramide signal amplification and RNA fluorescence in situ hybridization followed by immunofluorescence. The boundaries of the seminiferous tubules were detected with anti-ACTA2 (1:100, Santa Cruz, sc-32251), the staging of tubules was performed with anti-ACRV1 (1:200, Proteintech, 14040-1-AP) and Sertoli cell nuclei marked using anti-SOX9 (1:100, Sigma-Aldrich, HPA001758). See theSupplementary Methods for a detailed description of the experiments.
Mlh3−/− mice (The Jackson Laboratory; #018845) were bred at the Washington University in Saint Louis and processed as previously described (Jung et al., 2019 (link)). Mice of the Crispy line were bred by the Ahituv laboratory at the University of California, San Francisco, and fixed testes were received in 70% ethanol. Wild-type C57BL/6J (The Jackson Laboratory; #000664) were bred in-house at the Small Laboratory Animal Unit (ONPRC/OHSU). All animal experiments were performed in compliance with the regulations of the respective host institutions.
To capture subtle image differences resulting from experimental variables, the samples used to train the neural network were processed under different conditions: (i) fixation method—perfusion or immersion; (ii) fixative type—4% paraformaldehyde (PFA) or modified Davidson’s fixative and (iii) assay—IF with or without Tyramide signal amplification and RNA fluorescence in situ hybridization followed by immunofluorescence. The boundaries of the seminiferous tubules were detected with anti-ACTA2 (1:100, Santa Cruz, sc-32251), the staging of tubules was performed with anti-ACRV1 (1:200, Proteintech, 14040-1-AP) and Sertoli cell nuclei marked using anti-SOX9 (1:100, Sigma-Aldrich, HPA001758). See the
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