Organ Mapping Antibody Panels (OMAPs)

The goal of a published OMAP:

• Reduce development time and expense for researchers who want to do a similar experiment or panel.
• Standardize panels to increase data reproducibility while providing flexibility for research purposes.
• Provide a starting point for a new panel that is community-validated and peer reviewed.
• Support the development of a Human Reference Atlas.

OMAPs are community validated multiplex antibody panels that identify cell populations and structures within a diverse set of healthy tissues. This project originated from consortium members associated with the Human BioMolecular Atlas Program  (HuBMAP). Members within this group selected tissue specific antibodies and rigorously tested them for (1) high signal and low noise, (2) low non-specific interactions, (3) lower autofluorescence, and other attributes. Information about the antibodies is available on HuBMAP, publications, community databases, and/or vendor websites.

OMAPs are developed from tissues acquired from healthy or known disease-free organs. The information from these studies provides: (1) anatomical structure (2) cell types within these structures, (3) spatial location of cells, (4) intensity and size of cells and structures, (5) protein co-localization from conjugated antibody staining patterns.

OMAP information can be found on the OMAP HuBMAP CCF portal pages .

OMAPs can be used to design spatial biology experiments with antibodies used to label tissue samples. The design and validation of OMAPs allow researchers to confidently choose the antibody and reagents to generate spatial data without spending as much time re-validating panels or testing antibodies. OMAPs include information on tissue sample preparation, antigen retrieval, and staining methods. OMAP tables will include cell type markers and potential antibodies that maybe useful for the method used within the OMAP or other multiplexed imaging methods. Researchers may closely follow the panel design and method, or they can use the OMAP as an initial reference, to help guide the development of a multiplex panel for a particular tissue type.

OMAP tables link validated antibodies to cell types, providing a useful starting point for investigators seeking an antibody that will label their cell of interest in their tissue.

Researchers may closely follow the established panel or they can use the OMAP as an initial reference to guide development of a multiplexed imaging panel tailored to their questions of interest that is compatible with their sample preparation (FFPE, fixed frozen) and imaging method.

Examples of OMAPs used to understand tissue can be found in two publications included in the HuBMAP Nature Methods Collection .These publications used highly multiplexed imaging panels to evaluate the cellular neighborhoods and tissue organization of the human skin  and intestine .

Choose organ and multiplexed antibody–based imaging method

Click on the spreadsheet link (see red circle in the screenshot below) and download the file. Keep the information regarding how to cite the information for publications, grants, webpages, and other external documents.

The downloaded spreadsheet contains the information required to choose antibodies. It describes antibody name, clone, vendor, and conjugate type if applicable (e.g., fluorophore). Experimental design details include protocol method, tissue preservation method, and antibody dilution amount. In this example, the authors used IBEX , which is a multiplexed immunostaining protocol that employs iterative bleaching and cyclic antibody staining for high-parameter imaging. Cycle number, core panel, and essential 25-plex are important details, as they provide the sequence and grouping when using over 4 antibodies.

Table 1. Current OMAPs.

OMAP for multiplexed antibody-based imaging of human lymph node with IBEX

OMAP for multiplexed antibody-based imaging of human intestine with CODEX v1.0

OMAP for multiplexed antibody-based imaging of human skin with Cell DIVE v1.0

OMAP for multiplexed antibody-based imaging of human intestine with CODEX v1.0

OMAP for multiplexed antibody-based imaging of human intestine with CODEX v1.0

1. Quardokus EM, Saunders DC, McDonough E, Hickey JW, Werlein C, Surrette C, Rajbhandari P, Casals AM, Tian H, Lowery L, Neumann EK, Björklund F, Neelakantan TV, Croteau J, Wiblin AE, Fisher J, Livengood AJ, Dowell KG, Silverstein JC, Spraggins JM, Pryhuber GS, Deutsch G, Ginty F, Nolan GP, Melov S, Jonigk D, Caldwell MA, Vlachos IS, Muller W, Gehlenborg N, Stockwell BR, Lundberg E, Snyder MP, Germain RN, Camarillo JM, Kelleher NL, Börner K, Radtke AJ. Organ Mapping Antibody Panels: a community resource for standardized multiplexed tissue imaging. Nat Methods. 2023 Jul 19. PMID: 37468619. 
2. Radtke AJ, Quardokus EM, Saunders DC. SOP: Construction of Organ Mapping Antibody Panels for Multiplexed Antibody-Based Imaging of Human Tissues. DOI 10.5281/zenodo.5749882 
3. Hickey JW, Neumann EK, Radtke AJ, Camarillo JM, Beuschel RT, Albanese A, McDonough E, Hatler J, Wiblin AE, Fisher J, Croteau J, Small EC, Sood A, Caprioli RM, Angelo RM, Nolan GP, Chung K, Hewitt SM, Germain RN, Spraggins JM, Lundberg E, Snyder MP, Kelleher NL, Saka SK. Spatial mapping of protein composition and tissue organization: a primer for multiplexed antibody-based imaging. Nat Methods. 2022 Mar;19(3):284-295. PMID: 34811556. 
4. Radtke AJ, Chu CJ, Yaniv Z, Yao L, Marr J, Beuschel RT, Ichise H, Gola A, Kabat J, Lowekamp B, Speranza E, Croteau J, Thakur N, Jonigk D, Davis JL, Hernandez JM, Germain RN. IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues. Nat Protoc. 2022 Feb;17(2):378-401. PMID: 35022622. 
5. Du Z, Lin JR, Rashid R, Maliga Z, Wang S, Aster JC, Izar B, Sorger PK, Santagata S. Qualifying antibodies for image-based immune profiling and multiplexed tissue imaging. Nat Protoc. 2019 Oct;14(10):2900-2930. PMID: 31534232. 
6. Andrea J. Radtke. Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Lymph Node with IBEX.https://doi.org/10.48539/HBM624.PJSR.685 
7. Scalia CR, Boi G, Bolognesi MM, Riva L, Manzoni M, DeSmedt L, Bosisio FM, Ronchi S, Leone BE, Cattoretti G. Antigen Masking During Fixation and Embedding, Dissected. J Histochem Cytochem. 2017 Jan;65(1):5-20. Epub 2016 Oct 23. PMID: 27798289. 

1. HuBMAP CCF Portal: Organ Mapping Panels (OMAPs) 
2. Human Protein Atlas