OMAP-11: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Spleen with IBEX, v1.2

OMAP-11 was designed for IBEX imaging of fixed frozen human spleen samples (Radtke et al. 2022; 2020). The panel<br>includes 24 primary antibodies, 2 secondary antibodies, and the nuclear marker Hoechst for image alignment and<br>segmentation. It provides spatial context for the major anatomical structures and cell types featured in the ASCT+B<br>spleen table v1.3 (Jorgensen, Radtke, and Beuschel 2022), such as red pulp macrophages, splenic endothelial cells,<br>stromal cells, B cells, T cells, granulocytes, and platelets. This panel supports further phenotyping of B cells<br>(naïve, memory, switched), T cells (T follicular helper, regulatory), NK cells, dendritic cells (cDC1, cDC2,<br>plasmacytoid), and lymphatic endothelial cells with additional markers. Attempts to label CD271, a key splenic stromal<br>marker (Steiniger 2015), were unsuccessful using RRIDs AB_10982271 and AB_2564818 in this tissue context. Custom<br>antibodies were sourced from BioLegend or conjugated in-house using Abcam (ab269820) and Thermo Fisher (A20182,<br>A20188) kits. A representative dataset is available (Radtke et al. 2021), and further information on IBEX methods and<br>sample preparation is provided in the IBEX Knowledge-Base (Radtke et al. 2025; Yaniv et al. 2025).<br><br>**Bibliography:**<br><br>* Andrea J Radtke, Colin J Chu, Ziv Yaniv, Li Yao, James Marr, Rebecca T Beuschel, Hiroshi Ichise, et al. 2021. “Accompanying Dataset for: ‘IBEX: An Iterative Immunolabeling and Chemical Bleaching Method for High-Content Imaging of Diverse Tissues.’” Zenodo. https://zenodo.org/records/5244551.<br>* Jorgensen, Marda, Andrea J Radtke, and Rebecca T Beuschel. 2022. “Spleen (v1.3) Graph Data, v1.3.” 2022. https://lod.humanatlas.io/asct-b/spleen/v1.3/.<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., Evelyn Kandov, Bradley Lowekamp, Emily Speranza, Colin J. Chu, Anita Gola, Nishant Thakur, et al. 2020. “IBEX: A Versatile Multiplex Optical Imaging Approach for Deep Phenotyping and Spatial Analysis of Cells in Complex Tissues.” *Proceedings of the National Academy of Sciences* 117 (52): 33455–65. https://doi.org/10.1073/pnas.2018488117.<br>* Steiniger, Birte S. 2015. “Human Spleen Microanatomy: Why Mice Do Not Suffice.” *Immunology* 145 (3): 334–46. https://doi.org/10.1111/imm.12469.<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.