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These datasets were acquired using either the manual or automated IBEX multiplex imaging protocols and accompany the manuscript “IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues”, A. Radtke et al., 2021, Nature Protocols.
\n\nAll image data are stored using the Imaris file format. To view these multi-channel images, you can either use one of these free viewers, Imaris viewer, Fiji.
\n\nEach experiment has an associated imaging meta-data file in xlsx format and the resulting image in Imaris format.
\n\nHuman Jejunum (Automated)
\n\nDataset is a 24 parameter IBEX experiment performed on a human jejunum section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Thunder 3D Cell Culture microscope with a high precision (Quantum) stage, a high quantum efficiency sCMOS camera (DFC9000 GTC), and a 40x (1.3) NA oil objective. The light source was an LED8 with 8 individual LED lines for excitation with millisecond triggering. All images were captured at a 16-bit depth with the following pixel dimensions: x (0.160 μm), y (0.160 μm), and z (1 μm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.7.1.21655).
\n\nHuman Kidney (Automated)
\n\nDataset is a 16 parameter IBEX experiment performed on a human kidney Formalin-Fixed Paraffin-Embedded (FFPE) section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Thunder 3D Cell Culture microscope with a high precision (Quantum) stage, a high quantum efficiency sCMOS camera (DFC9000 GTC), and a 40x (1.3) NA oil objective. The light source was an LED8 with 8 individual LED lines for excitation with millisecond triggering. All images were captured at a 16-bit depth with the following pixel dimensions: x (0.160 μm), y (0.160 μm), and z (1 μm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.7.1.21655).
\n\nHuman Lymph Node (Automated)
\n\nDataset is a 25 parameter IBEX experiment performed on a human lymph node section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Thunder 3D Cell Culture microscope with a high precision (Quantum) stage, a high quantum efficiency sCMOS camera (DFC9000 GTC), and a 40x (1.3) NA oil objective. The light source was an LED8 with 8 individual LED lines for excitation with millisecond triggering. All images were captured at a 16-bit depth with the following pixel dimensions: x (0.160 μm), y (0.160 μm), and z (1 μm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.7.1.21655).
\n\nHuman Skin (Automated)
\n\nDataset is an 18 parameter IBEX experiment performed on a human skin section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Thunder 3D Cell Culture microscope with a high precision (Quantum) stage, a high quantum efficiency sCMOS camera (DFC9000 GTC), and a 40x (1.3) NA oil objective. The light source was an LED8 with 8 individual LED lines for excitation with millisecond triggering. All images were captured at a 16-bit depth with the following pixel dimensions: x (0.160 μm), y (0.160 μm), and z (1 μm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.7.1.21655).
\n\nHuman Liver (Manual)
\n\nDataset is a 22 parameter IBEX experiment performed on a human liver section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Leica TCS SP8 X confocal microscope equipped with a 40X objective (NA 1.3), 4 HyD and 1 PMT detectors, a white light laser that produces a continuous spectral output between 470 and 670 nm as well as 405, 685, and 730 nm lasers. All images were captured at an 8-bit depth, with a line average of 3, and 1024x1024 format with the following pixel dimensions: x (0.284 µm), y (0.284 µm), and z (1 µm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.5.5.19976).
\n\nHuman Lymph Node (Manual)
\n\nDataset is a 38 parameter IBEX experiment performed on a human lymph node section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Leica TCS SP8 X confocal microscope equipped with a 40X objective (NA 1.3), 4 HyD and 1 PMT detectors, a white light laser that produces a continuous spectral output between 470 and 670 nm as well as 405, 685, and 730 nm lasers. All images were captured at an 8-bit depth, with a line average of 3, and 1024x1024 format with the following pixel dimensions: x (0.284 µm), y (0.284 µm), and z (1 µm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.5.5.19976).
\n\nHuman Spleen (Manual)
\n\nDataset is a 25 parameter IBEX experiment performed on a human spleen section labeled with the nuclear marker Hoechst and antibodies directed against the indicated markers. Images were acquired using an inverted Leica TCS SP8 X confocal microscope equipped with a 40X objective (NA 1.3), 4 HyD and 1 PMT detectors, a white light laser that produces a continuous spectral output between 470 and 670 nm as well as 405, 685, and 730 nm lasers. All images were captured at an 8-bit depth, with a line average of 3, and 1024x1024 format with the following pixel dimensions: x (0.284 µm), y (0.284 µm), and z (1 µm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.5.5.19976).
", "languages": [ { "id": "eng", "title": { "en": "English" } } ], "publication_date": "2021-08-24", "publisher": "Zenodo", "resource_type": { "id": "dataset", "title": { "de": "Datensatz", "en": "Dataset" } }, "rights": [ { "description": { "en": "The Creative Commons Attribution license allows re-distribution and re-use of a licensed work on the condition that the creator is appropriately credited." }, "icon": "cc-by-icon", "id": "cc-by-4.0", "props": { "scheme": "spdx", "url": "https://creativecommons.org/licenses/by/4.0/legalcode" }, "title": { "en": "Creative Commons Attribution 4.0 International" } } ], "subjects": [ { "subject": "Multiplexed antibody-based imaging, tissue systems biology, serial immunofluorescence microscopy, single-cell analysis" } ], "title": "Accompanying dataset for: \"IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues\"" }, "parent": { "access": { "owned_by": { "user": 165393 } }, "communities": { "default": "9256ae8c-7b47-4ce5-a8ac-23f7d2a6ca77", "entries": [ { "access": { "member_policy": "open", "members_visibility": "public", "record_policy": "open", "review_policy": "open", "visibility": "public" }, "children": { "allow": false }, "created": "2023-05-09T23:56:38.617371+00:00", "custom_fields": {}, "deletion_status": { "is_deleted": false, "status": "P" }, "id": "9256ae8c-7b47-4ce5-a8ac-23f7d2a6ca77", "links": {}, "metadata": { "curation_policy": "Uploads are accepted from OMAP authors. For information about authoring an OMAP, please see SOP: Construction of Organ Mapping Antibody Panels for Multiplexed Antibody-Based Imaging of Human Tissues.
\r\n", "page": "Organ Mapping Antibody Panels (OMAPs) are collections of antibodies that allow spatial mapping of the anatomical structures and cell types present in diverse organs using multiplexed antibody-based imaging.
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