{"id": "package:7765fa05-1485-4671-9a8c-7a896e6ff24c", "name": "171_Adult_Left_Colon_4689_20x_anti_HucD_594_anti_calretinin_647_1.tif", "self_uri": "https://services.scicrunch.io/sparc/drs/v1/objects/7765fa05-1485-4671-9a8c-7a896e6ff24c", "size": 95103447, "created_time": "2019-07-10T05:05:36,331205Z", "updated_time": "2022-12-13T15:43:03,032359Z", "version": "1", "mime_type": "image/tiff", "checksums": [{"checksum": "1704c29d12d9c11a5406c9d8a56c329e5e187936601c427d5429bea3947eafe1", "type": "sha256"}], "access_methods": [{"type": "s3", "access_url": {"url": "s3://prd-sparc-discover50-use1/55/files/primary/Human Colon 3D/sub_4689/171_Adult_Left_Colon_4689_20x_anti_HucD_594_anti_calretinin_647_1.tif"}, "region": "us-east-1"}], "dataset": {"id": "55", "doi": "DOI:10.26275/pzek-91wx", "title": "Robust 3-Dimensional visualization of human colon enteric nervous system without tissue sectioning", "description": "Details a new technique to image the enteric nervous system (ENS) and other cells that control motility in the human colon in three dimensions, by combining tissue clearing, immunohistochemistry, confocal microscopy, and quantitative image analysis.", "abstract": "Background & Aims: Current understanding and management of bowel motility disorders is limited by histologic techniques that rarely yield valuable information about cells that control motility. These cells are deep in the bowel wall, not uniform in distribution, and impossible to see in sufficient detail in small two dimensional sections routinely used for human pathology. Methods: We developed a new technique to image in three dimensions the enteric nervous system (ENS) and other cells that control motility in human colon by combining tissue clearing, immunohistochemistry, confocal microscopy, and quantitative image analysis. Our methods do not require tissue sectioning and can be scaled to evaluate large bowel regions. Results: Using human adult colon, one of the thickest bowel regions, we tested many approaches and dozens of antibodies until we established a method that worked consistently to visualize the human ENS in three dimensions. By imaging colon from people who do not have bowel motility disorders, we generated unprecedented images, and we provide detailed quantitative data about the density of neurons and glia, ratios of cholinergic and nitrergic neurons, and organization of the ENS. Murine images are shown for comparison. Conclusions: Most of what we need to know about ENS structure is only visible in three dimensions. Our new techniques and quantitative data provide the opportunity to radically change how we evaluate bowel dysmotility and may be applicable to other types of bowel disease."}}