Near-isotropic, aligned volume electron microscopy (eFIB-SEM) of Mouse hippocampus (jrc_mus-hippocampus-2)

Sample: 2–3 month old male mouse, strain: C57BL/6J from Jackson Lab.
Sample Description: This dataset was acquired from the CA1 region of the mouse hippocampus using a hybrid fixation and staining protocol optimized for primary cilia visualization. Tissue from a 2–3-month-old male C57BL/6J mouse was perfused with 3% paraformaldehyde, sectioned, and processed through high-pressure freezing in 1-hexadecene. The sample was freeze-substituted in osmium tetroxide, uranyl acetate, and either 3-amino-1,2,4-triazole or imidazole, and embedded in Durcupan resin. Sample preparation was performed by Shu-Hsien Sheu. Imaging and post-processing were completed by C. Shan Xu (Yale), Gleb Shtengel, and Harald Hess (HHMI/Janelia).
Protocol: Two datasets were acquired from the CA1 region of the mouse hippocampus using a hybrid fixation and staining protocol optimized for primary cilia visualization. 2–3-month-old male C57BL/6 mice were perfused with 3% paraformaldehyde and post-fixed in modified PFA buffer for 2 hours. Tissue was rinsed in 400 mOsM buffer and sectioned (100 µm) on a vibratome. Sections were fixed overnight in 1% PFA and 2% glutaraldehyde. 2 mm punches were high-pressure frozen in 1-hexadecene, freeze-substituted in osmium tetroxide, uranyl acetate, and either 3-amino-1,2,4-triazole or imidazole. Samples were embedded in Durcupan resin.
Contributions: Sample prepared by Shu-Hsien Sheu. Imaging and post-processing by C. Shan Xu (Yale), Gleb Shtengel, and Harald Hess(HHMI/Janelia).
Acquisition ID: jrc_mus-hippocampus-2
Voxel size (nm): 8 x 8 x 8 (x, y, z)
Data dimensions (µm): 44.98 x 44.9 x 55.06 (x, y, z)
Imaging start date: 2017-10-04
Imaging duration (days): 8
Landing energy (eV): 1100
Imaging current (nA): 1
Scanning speed (MHz): 1.250
Dataset URL: s3://janelia-cosem-datasets/jrc_mus-hippocampus-2/jrc_mus-hippocampus-2.zarr/recon-1/em/
Visualization Website: https://openorganelle.janelia.org/datasets/jrc_mus-hippocampus-2
Publication: Sheu, et al., 2022; Xu et al., 2017

样本：2~3月龄雄性小鼠，品系为C57BL/6J，购自杰克逊实验室（Jackson Lab）。
样本描述：本数据集采集自小鼠海马CA1区，采用经优化的、用于可视化初级纤毛（primary cilia）的混合固定与染色方案。实验取材自2~3月龄雄性C57BL/6J小鼠的脑组织，经3%多聚甲醛灌注固定后切片，随后在1-十六烯中完成高压冷冻。样本经四氧化锇、醋酸铀及3-氨基-1,2,4-三唑或咪唑进行冷冻替代，之后包埋于Durcupan树脂中。样本制备由Shu-Hsien Sheu完成，成像与后处理由耶鲁大学的C. Shan Xu、Gleb Shtengel以及HHMI/Janelia的Harald Hess共同完成。
实验方案：本数据集包含两组从小鼠海马CA1区采集的数据，采用经优化的、用于可视化初级纤毛（primary cilia）的混合固定与染色方案。实验使用2~3月龄雄性C57BL/6小鼠，经3%多聚甲醛灌注后，在改良PFA缓冲液中后固定2小时。脑组织经400 mOsM缓冲液漂洗后，通过振动切片机切成100 µm厚的组织切片。切片于1%多聚甲醛与2%戊二醛中固定过夜。切取2 mm直径的组织块，在1-十六烯中完成高压冷冻，经四氧化锇、醋酸铀及3-氨基-1,2,4-三唑或咪唑进行冷冻替代后，包埋于Durcupan树脂中。
贡献：样本制备由Shu-Hsien Sheu完成。成像与后处理由耶鲁大学的C. Shan Xu、Gleb Shtengel以及HHMI/Janelia的Harald Hess共同完成。
采集ID：jrc_mus-hippocampus-2
体素尺寸（纳米）：8 × 8 × 8（x, y, z轴）
数据维度（微米）：44.98 × 44.9 × 55.06（x, y, z轴）
成像起始日期：2017-10-04
成像时长（天）：8
着陆能量（电子伏特）：1100
成像电流（纳安）：1
扫描速度（兆赫兹）：1.250
数据集链接：s3://janelia-cosem-datasets/jrc_mus-hippocampus-2/jrc_mus-hippocampus-2.zarr/recon-1/em/
可视化网站：https://openorganelle.janelia.org/datasets/jrc_mus-hippocampus-2
相关文献：Sheu等人，2022年；Xu等人，2017年