Near-isotropic, aligned volume electron microscopy (eFIB-SEM) of Mouse deep nuclei (jrc_mus-cerebellum-5)

Sample: Wild-type, 8 month old mouse, strain: C57BL/6;129 from Gene Targeting & Transgenic Facility of HHMI Janelia Research CampusSample Description: The mouse deep cerebellar nuclei are vital components of the cerebellum that help regulate motor control, balance, and motor learning. They integrate inputs from the cerebellar cortex and spinal cord, sending output to other brain regions to coordinate movement. Studies using electron microscopy have advanced our understanding of the synaptic organization and neurotransmitter dynamics in these nuclei, shedding light on how the cerebellum adapts to learning and how dysfunction in these areas leads to motor disorders. The mouse model is particularly valuable for studying cerebellar function and its role in various neurodegenerative and neurodevelopmental disorders.Protocol: 8-month-old female C57BL6/129 mice were anesthetized with a ketamine/xylazine anesthetic mixture before perfusion with 2% depolymerized paraformaldehyde and 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer. Samples were postfixed in 2% OsO₄, 1.5% K₄Fe(CN)₆, and 0.1 M sodium cacodylate buffer for 1 hour. Subsequently, specimens were stained en bloc with 2% aqueous uranyl acetate (1 hour), dehydrated in increasing concentrations of ethanol and propylene oxide, embedded in Epon, and then re-embedded in Durcupan.Contributions: Sample provided by Yumei Wu, Christina Whites, and Pietro De Camilli (Yale), prepared for imaging by Yumei Wu and Christina Whites (Yale), with imaging and post-processing by Wei Qui, Gleb Shtengel, and Harald Hess(HHMI/Janelia).Acquisition ID: jrc_mus-cerebellum-5Voxel size (nm): 4 x 4 x 4 (x, y, z)Data dimensions (µm): 20.57 x 20.46 x 11.6 (x, y, z)Imaging start date: 2023-09-25Imaging duration (days): 22Primary energy (eV): 900Imaging current (nA): 0.25Scanning speed (MHz): 0.100Dataset URL: https://open.quiltdata.com/b/janelia-cosem-datasets/tree/jrc_mus-cerebellum-5/jrc_mus-cerebellum-5.zarr/recon-1/em/Visualization Website: https://openorganelle.janelia.org/datasets/jrc_mus-cerebellum-5Publication: Xu et al., 2020; Xu et al, 2021