OMAP-12: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Retina with IBEX, v1.2

OMAP-12 was developed for IBEX (Iterative Bleaching Extends multi-pleXity) imaging of fixed frozen human retina<br>samples (Radtke et al. 2022; 2020). The panel includes 14 antibodies, 3 lectins, and DAPI for nuclear segmentation and<br>image alignment. It captures spatial information for key anatomical structures and cell types in the healthy human<br>retina, such as photoreceptors, bipolar cells, ganglion cells, astrocytes, microglia, Muller glia, and retinal<br>vascular endothelial cells. While major neuronal classes are visualized, subtypes (e.g., amacrine, ganglion, and<br>bipolar cell subtypes) are not individually distinguished due to reagent limitations. Fluorescent lectins PNA,<br>Isolectin B4, and Phalloidin were used to label cone photoreceptors, vascular endothelium, and the outer limiting<br>membrane, respectively, but are not part of the antibody panel and thus excluded from the OMAP antibody table. Some<br>antibodies were custom conjugated per manufacturer protocols. A representative dataset generated with this OMAP is<br>available (Chu 2023). More information on IBEX sample preparation and imaging techniques is provided in the IBEX<br>Knowledge-Base (Radtke et al. 2025; Yaniv et al. 2025).<br><br>**Bibliography:**<br><br>* Chu, Colin J. 2023. “Human Retina OMAP-12.” Zenodo. https://doi.org/10.5281/zenodo.7777286.<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>* 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 Bio* 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.