OMAP-19: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Palatine Tonsil with IBEX, v1.0

OMAP-19 was designed for IBEX (Iterative Bleaching Extends multi-pleXity) imaging of human FFPE tonsil samples<br>sectioned onto glass slides (https://doi.org/10.1038/s41596-021-00644-9). The panel consists of 6 cycles of 20 primary<br>antibodies, 5 secondary antibodies, and the nuclear label Hoechst for image alignment and nuclear segmentation. This<br>OMAP provides a spatial context for 10 anatomical structures and at least 16 cell types present in the ASCT+B tonsil<br>table. OMAP-19 was designed to examine myeloid subsets (CD11b, CD11c, CD14, CD15, HLA-DR) and angiogenesis (VEGF-A and<br>VEGF-C) in human ovarian cancer biopsies. The human tonsil was used as a positive control for antibody validation and<br>panel development. OMAP-19 has 11 markers in common with the tonsil MICS panel (OMAP-10,<br>https://doi.org/10.48539/HBM663.JJMJ.628); however, there is no overlap between the antibodies used across the tonsil<br>OMAPs. This panel does not allow for visualization of the mantle zone, naive B cells, plasma cells, or cycling B cells<br>enriched in the dark zone of the germinal center. Prior to immunolabeling, antigen retrieval was performed using a<br>pressure cooker (Borg Decloaker BD1000, 110°C for 15 minutes) and a tris-based buffer (pH 9.5). Several custom<br>antibodies were created using commercial labeling kits as indicated in the associated RRID entries for these<br>antibodies (AB_3095771, RRID:AB_3095773, and RRID:AB_3095774). More details on the antigen retrieval protocol and<br>step-by-step application of antibodies can be found on the [IBEX Imaging Community<br>site](https://ibeximagingcommunity.github.io/ibex_imaging_knowledge_base/). A representative dataset and associated<br>channel metadata has been uploaded to Zenodo (https://zenodo.org/records/11222093).