High-speed mapping of whole-mouse peripheral nerves at subcellular resolution

In contrast to the rapid advancements in mesoscale connectomic mapping of the mammalian brain, similar mapping the peripheral nervous system has remained challenging due to the body's size and complexity. Here, we present a high-speed blockface volumetric imaging system (blockface-VISoR) with an optimized workflow of whole-body clearing. This innovative system enables imaging of the entire adult mouse (~ 4 × 2 × 8 cm³) at a voxel resolution of 1 × 1 × 2.5 μm³ within 40 hours.Here are some demo datasets imaged by blockface-VISoR system, including raw slice images, downsampled reconstructed whole-body images and reconstructed regions of interest (ROIs). For ease of data transfer, certain individual files are divided into smaller compressed files. Users can extract and reassemble these files using 7-Zip. 1.     Thy1_origin_Data: Contains demo raw volumetric images from a mouse body for validating the blockface-VISoR imaging technique. Users can open the Raw.ome.tiff file in each slice fold using ImageJ/Fiji.2.     Thy1_Reconstruction_4μm: Provides 4× downsampled image sequences (at a resolution of 4 × 4 × 4 μm3) of a whole Thy1-EGFP transgenic mouse labeled with tomato lectin conjugated with DyLight 649. The images are compatible with ImageJ/Fiji or can be converted to Imaris format using ImarisFileConverter for viewing in Imaris Viewer or Lychnis.3.     Sympathetic ROI: Contains downsampled ROIs of a whole mouse with sympathetic nerves labeling. These .ims format files are viewable in Imaris Viewer or Lychnis.4.     Vagal innervations: is an ROI of a whole mouse with vagus nerves labeling. This file can be opened by Imaris Viewer or Lychnis. The complete uncompressed raw image datasets of whole mouse are too large (exceeding 70 terabytes per channel) for public data repositories. A free and more convenient view of the whole-mouse images are publicly accessible through our website (https://smart.siat.ac.cn/meso-example.html). Additionally, other datasets involved in our published work are available to all scientists upon request to the lead contact by feasible data transferring methods including cloud storage, physical hard disk drives, or onsite visiting.