{"id": "package:ebbc8524-5f29-4811-b910-58bd80c45ff6", "name": "20171122P_pC_M_PGP9.5_DAPI_DIVESP8_10x_5.jp2", "self_uri": "https://services.scicrunch.io/sparc/drs/v1/objects/ebbc8524-5f29-4811-b910-58bd80c45ff6", "size": 258930, "created_time": "2023-04-13T18:49:07,771203Z", "updated_time": "2023-04-13T18:54:44,729322Z", "version": "1", "mime_type": "image/jp2", "checksums": [{"checksum": "c7ed0e35d3b3e64f308d46805cafffa6", "type": "sha256"}], "access_methods": [{"type": "s3", "access_url": {"url": "s3://prd-sparc-discover50-use1/331/files/derivative/sub-20171122P/sam-20171122P-pC/20171122P_pC_M_PGP9.5_DAPI_DIVESP8_10x_5.jp2"}, "region": "us-east-1"}], "dataset": {"id": "331", "doi": "DOI:10.26275/rhda-nblc", "title": "CLARITY and 3D imaging with high resolution and deep scanning of innervation in the pig colon by using SP8 DIVE fully tunable spectral multiphoton microscope", "description": "Deep scanning of innervation in the pig colon by using SP8 DIVE multiphoton microscope", "abstract": "To demonstrate the innervation of the pig proximal colon mucosa in 3D structure with high resolution by deep scanning using SP8 DIVE multiphoton microscope, a latest multiphoton microscope with fully spectrally tunable detection. The pig proximal colon samples were cleared by a modified passive CLARITY protocol we developed in first SPARC funding year (https://dx.doi.org/10.17504/protocols.io.4r9gv96). Immunofluorescent methods were used to label the nerve fibers in the mucosa by using antibodies against PGP9.5, Neurofilement (NF) and vasoactive intestinal polypeptide (VIP).  Images were acquired with Leica TCS SP8 DIVE multiphoton microscope (Leica Microsystem). The Z-axis intervals were 1 µm with depths between 500 – 1000 µm. The images were collected and processed for 3D reconstruction and video generation using Leica Application Suite software (LAS X 3.0, Leica Microsystem) and Imaris 9.2 for Neuroscientists (Bitplane Inc., Concord, MA).  The projections of these fine fibers from inner submucosal neurons and sprouting into mucosa with multiple branches were also clearly displayed by videos in a 360° horizontal or vertical rotation. The dataset of 3D images and videos of innervation in the pig colonic mucosa obtained by using CLARITY/SP8 DIVE multiphoton microscope with high resolution and deep scanning provides foundermental information to distinguish the different classes of nerve fibers, visualize their projections to different layers and compare their differences among the segments, which are not readily revealed by conventional confocal microscope and rarely reported before."}}