OMAP-19: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Palatine Tonsil with IBEX, v1.1

OMAP-19 was designed for IBEX (Iterative Bleaching Extends multi-pleXity) imaging of human FFPE tonsil tissue sections<br>on glass slides (Radtke et al. 2022). The panel comprises 6 imaging cycles with 20 primary antibodies, 5 secondary<br>antibodies, and the nuclear label Hoechst for image registration and segmentation. It provides spatial mapping for 10<br>anatomical structures and at least 16 cell types included in the ASCT+B tonsil table. Though initially developed to<br>study myeloid subsets and angiogenesis markers in human ovarian cancer biopsies, OMAP-19 uses tonsil tissue as a<br>positive control for antibody validation and panel optimization. The myeloid targets include CD11b, CD11c, CD14, CD15,<br>and HLA-DR, while VEGF-A and VEGF-C were selected for vascular profiling. OMAP-19 shares 11 markers with OMAP-10<br>(Tonsil MICS), but employs different antibody clones. This panel does not support visualization of the mantle zone,<br>naïve B cells, plasma cells, or dark zone B cells of the germinal center. Antigen retrieval was conducted using a<br>pressure cooker (Borg Decloaker BD1000) at 110°C for 15 minutes in a tris-based buffer (pH 9.5). Several antibodies<br>were custom-labeled using commercial kits, with corresponding RRIDs (AB_3095771, AB_3095773, AB_3095774) provided. A<br>full protocol and antibody application guide is available on the [IBEX Imaging Community<br>site](https://ibeximagingcommunity.github.io/ibex_imaging_knowledge_base/). Representative datasets and channel<br>metadata have been uploaded to Zenodo (Padhan et al. 2024). Additional technical details, image analysis workflows,<br>and community resources are documented in the IBEX Knowledge-Base (Radtke et al. 2025; Yaniv et al. 2025).<br><br>**Bibliography:**<br><br>* Müller, Werner, Silvia Rüberg, and Andreas Bosio. 2023. “10-Palatine-Tonsil-Mics (v1.0) Graph Data, v1.0.” https://lod.humanatlas.io/omap/10-palatine-tonsil-mics/v1.0/.<br>* Padhan, Kartika, Andrea J. Radtke, Elena Giudice, Ronald N. Germain, and Jung-Min Lee. 2024. “OMAP-19 Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Tonsil with IBEX.” Zenodo. https://zenodo.org/records/11222093.<br>* Radtke, Andrea J., Colin J. Chu, Ziv Yaniv, Li Yao, James Marr, Rebecca T. Beuschel, Hiroshi Ichise, et al. 2022. “IBEX: An Iterative Immunolabeling and Chemical Bleaching Method for High-Content Imaging of Diverse Tissues.” *Nature Protocols* 17 (2): 378–401. https://doi.org/10.1038/s41596-021-00644-9.<br>* Radtke, Andrea J., Ifeanyichukwu U. Anidi, Leanne Arakkal, Armando J. Arroyo-Mejías, Rebecca T. Beuschel, Katy Börner, Colin J. Chu, et al. 2025. “The IBEX Knowledge-Base: A Central Resource for Multiplexed Imaging Techniques.” *PLoS Biol* 23(3): e3003070. https://doi.org/10.1371/journal.pbio.3003070.<br>* Yaniv, Ziv, Ifeanyichukwu U. Anidi, Leanne Arakkal, Armando J. Arroyo-Mejías, Rebecca T. Beuschel, Katy Börner, Colin J. Chu, et al. 2025. “The IBEX Imaging Knowledge-Base: A Community Resource Enabling Adoption and Development of Immunofluorescence Imaging Methods.” *eLife* 14:RP105737.