| UniProt Accession Number | Reagent Type | Target Name / Protein Biomarker | Target Species | Host Organism | Isotype | Clonality | Vendor | Catalog Number | Conjugate | RRID | Availability | Method | Tissue Preservation | Target Tissue | Tissue State | Detergent | Antigen Retrieval Conditions | Dye Inactivation Conditions | Recommend | Agree | Disagree | Contributor | Notes |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| NA | Nuclear Dye | Hoechst 33342 | Human | NA | NA | NA | Biotium | 40046 | NA | NA | Stock | IBEX2D Automated | 1:4 Cytofix/Cytoperm Fixed Frozen | Jejunum | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [1] | NA | 0000-0003-4379-8967 | 1 |
| NA | Nuclear Dye | Hoechst 33342 | Human | NA | NA | NA | Biotium | 40046 | NA | NA | Stock | IBEX2D Automated | 1:4 Cytofix/Cytoperm Fixed Frozen | Lymph Node | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [1] | NA | 0000-0003-4379-8967 | 1 |
| NA | Nuclear Dye | Hoechst 33342 | Human | NA | NA | NA | Biotium | 40046 | NA | NA | Stock | IBEX2D Automated | FFPE | Kidney | NA | 0.3% Triton-X-100 | pH 6 for 40 minutes at 95C (AR6 Akoya Biosciences AR600250ML) | NA | Yes | 0000-0003-4379-8967 [1] | NA | 0000-0003-4379-8967 | 1 |
| NA | Nuclear Dye | Hoechst 33342 | Human | NA | NA | NA | Biotium | 40046 | NA | NA | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Spleen | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [1, 2] | NA | 0000-0003-4379-8967 | 1 |
| NA | Nuclear Dye | Hoechst 33342 | Human | NA | NA | NA | Biotium | 40046 | NA | NA | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Lymph Node | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [1, 2] | NA | 0000-0003-4379-8967 | 1 |
Publications
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A. J. Radtke et al., “IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues”, Nat. Protoc., 17(2):378-401, 2022, doi: 10.1038/s41596-021-00644-9.
A. J. Radtke et al., “Accompanying dataset for: IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues”, doi: 10.5281/zenodo.5244550.
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A. J. Radtke et al., “IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues”, Proc Natl Acad Sci, 117(52):33455–33465, 2020, doi:10.1073/pnas.2018488117
Additional Notes
- Does not bleach. Use as a fiducial for image alignment and object-based segmentation.
| Human kidney: Hoechst (blue, catalog number 40046) and Vimentin (magenta, catalog number 677807) |
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| Human jejunum: Hoechst (blue, catalog number 40046), CD106 (cyan, catalog number 305806), and CD31 (red, catalog number 303106) |
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| Human lymph node: Hoechst (blue, catalog number 40046) and Collagen IV (red, catalog number ab6586) |
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