Segmentation of isotropic 3D electron microscopy (FIB-SEM) data of mouse guard hair follicle (jrc_mus-guard-hair-follicle)

<b>Sample:</b> Guard hair follicle from the back of a 3-week-old C57BL/6J WT mouse<b>Sample Description:</b> Across mammalian skin, structurally complex and diverse mechanosensory end organs respond to mechanical stimuli and enable our perception of dynamic, light touch. How forces act on morphologically dissimilar mechanosensory end organs of the skin to gate the requisite mechanotransduction channel Piezo2 and excite mechanosensory neurons is not understood. Here, we report high-resolution reconstructions of the hair follicle lanceolate complex, Meissner corpuscle, and Pacinian corpuscle and the subcellular distribution of Piezo2 within them. Across all three end organs, Piezo2 is restricted to the sensory axon membrane, including axon protrusions that extend from the axon body. These protrusions, which are numerous and elaborate extensively within the end organs, tether the axon to resident non-neuronal cells via adherens junctions. These findings support a unified model for dynamic touch in which mechanical stimuli stretch hundreds to thousands of axon protrusions across an end organ, opening proximal, axonal Piezo2 channels and exciting the neuron.This dataset contains manually proofread automatic segmentation of the FIB-SEM dataset in jrc_mus-guard-hair-follicle.<b>Protocol:</b> Samples were dissected and drop fixed in glutaraldehyde and paraformaldehyde, and then osmicated with osmium tetroxide and potassium ferrocyanide, followed by osmium tetroxide only. Samples were subsequently stained with uranyl acetate and samarium chloride. Samples were dehydrated with an ethanol series followed by anhydrous acetone, infiltrated with Durcupan resin, and cured at 60°C.<b>Contributions:</b> Sample provided by Annie Handler (Harvard Medical School/HHMI) and Qiyu Zhang (Harvard Medical School/HHMI), prepared for imaging by Song Pang (HHMI/Janelia, currently at Yale School of Medicine), imaged by Song Pang and C. Shan Xu (HHMI/Janelia, currently at Yale School of Medicine), post data registration by C. Shan Xu, global image alignment and processing by Annie Handler and Qiyu Zhang, automatic segmentation by Tri M. Nguyen (Harvard Medical School) under the supervision of Wei-Chung Allen Lee (Harvard Medical School), ground truth annotation by Rebecca Plumb, Brianna Sanchez, Karyl Ashjian, Aria Shotland, Bartianna Brown, Madiha Kabeer, Nusrat Africawala, Stuart Cattel, Annie Handler, and Qiyu Zhang (all Harvard Medical School/HHMI), and segmentation proofreading by Annie Handler, Qiyu Zhang, and Michael Nolan-Tamariz (Harvard Medical School/HHMI).<b>Acquisition ID:</b> jrc_mus-guard-hair-follicle<b>Voxel size (nm):</b> 6 x 6 x 6 (x, y, z)<b>Data dimensions (µm): </b>88.1 x 95.9 x 77.6 (x, y, z)<b>Scanning speed (MHz): </b>1<b>Dataset URL: </b>s3://janelia-cosem-datasets/jrc_mus-guard-hair-follicle/jrc_mus-guard-hair-follicle.zarr/recon-1/labels/inference/segmentations/<b>EM DOI:</b> https://doi.org/10.25378/janelia.23969052<b>Visualization Website</b>: https://openorganelle.janelia.org/datasets/jrc_mus-guard-hair-follicle<b>Publication: </b>Handler, et al., 2023<br>

样本：3周龄C57BL/6J野生型（WT）小鼠背部的护毛囊
样本描述：在哺乳动物皮肤中，结构复杂且形态多样的机械感觉终末器官可响应机械刺激，使我们能够感知动态轻触。目前尚不明确，作用力如何作用于皮肤中形态各异的机械感觉终末器官，以门控必需的机械转导通道Piezo2并激活机械感觉神经元。本研究报道了毛囊柳叶状神经末梢复合体（lanceolate complex）、迈斯纳小体（Meissner corpuscle）与帕奇尼小体（Pacinian corpuscle）的高分辨率重构结果，以及Piezo2在这些结构中的亚细胞分布。在这三种终末器官中，Piezo2均局限于感觉轴突膜，包括从轴突本体延伸出的轴突突起。这些突起数量众多，在终末器官内广泛延展，通过黏着连接将轴突锚定在驻留的非神经元细胞上。上述研究结果支持一种动态触觉的统一模型：机械刺激拉伸终末器官内成百上千的轴突突起，从而激活轴突近端的Piezo2通道并兴奋神经元。本数据集包含针对jrc_mus-guard-hair-follicle的聚焦离子束扫描电子显微镜（FIB-SEM）数据集进行人工校对后的自动分割结果。
实验流程：样本经解剖后，于戊二醛与多聚甲醛中滴加固定，随后使用四氧化锇与铁氰化钾进行锇化处理，再单独使用四氧化锇处理。后续样本依次经醋酸铀酰与氯化钐染色，通过梯度乙醇系列脱水，再经无水丙酮置换，用Durcupan树脂渗透后，于60℃下固化。
贡献方：样本由Annie Handler（哈佛医学院/霍华德·休斯医学研究所，HHMI）与Qiyu Zhang（哈佛医学院/HHMI）提供，成像前样本制备由Song Pang（HHMI/简维亚研究所，现任职于耶鲁医学院）完成，成像由Song Pang与C. Shan Xu（HHMI/简维亚研究所，现任职于耶鲁医学院）执行，数据配准后处理由C. Shan Xu完成，全局图像配准与处理由Annie Handler与Qiyu Zhang完成，自动分割由Tri M. Nguyen（哈佛医学院）在Wei-Chung Allen Lee（哈佛医学院）的指导下完成，真值标注由Rebecca Plumb、Brianna Sanchez、Karyl Ashjian、Aria Shotland、Bartianna Brown、Madiha Kabeer、Nusrat Africawala、Stuart Cattel、Annie Handler与Qiyu Zhang（均任职于哈佛医学院/HHMI）完成，分割结果校对由Annie Handler、Qiyu Zhang与Michael Nolan-Tamariz（哈佛医学院/HHMI）完成。
采集ID：jrc_mus-guard-hair-follicle
体素尺寸（nm）：6 × 6 × 6（x、y、z轴）
数据尺寸（µm）：88.1 × 95.9 × 77.6（x、y、z轴）
扫描速度（MHz）：1
数据集链接：s3://janelia-cosem-datasets/jrc_mus-guard-hair-follicle/jrc_mus-guard-hair-follicle.zarr/recon-1/labels/inference/segmentations/
电子显微镜数据集DOI：https://doi.org/10.25378/janelia.23969052
可视化网站：https://openorganelle.janelia.org/datasets/jrc_mus-guard-hair-follicle
相关论文：Handler等人，2023年