OMAP-11: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Spleen with IBEX, v1.1

OMAP-11 was designed for IBEX imaging of fixed frozen human spleen samples<br>(https://doi.org/10.1038/s41596-021-00644-9; https://doi.org/10.1073/pnas.2018488117). The panel contains 24 primary<br>antibodies, 2 secondary antibodies, and Hoechst for image alignment and nuclear segmentation. This OMAP provides a<br>spatial context for the main anatomical structures and cell types present in the ASCT+B spleen table v1.3 including<br>red pulp macrophages, splenic endothelial cells, stromal cells of the red and white pulp, B cells, T cells,<br>granulocytes, and platelets. This panel can be expanded to include more granular phenotyping of B cells (naïve,<br>memory, switched), T cells (T follicular helper cells, T regulatory cells), NK cells, dendritic cells (cDC1, cDC2,<br>plasmacytoid DCs), and lymphatic endothelial cells with the addition of appropriate markers. Several antibodies for<br>CD271, a critical splenic stromal marker (https://doi.org/10.1111/imm.12469), were tested but did not provide reliable<br>labeling in the spleen for the tissue preservation method employed for this study<br>(RRIDs AB_10982271, AB_2564818). Custom antibodies were purchased from BioLegend or conjugated in house<br>using labeling kits from Abcam (ab269820) and Thermo Fisher Scientific (A20182, A20188). A representative<br>dataset acquired using this OMAP can be found here: https://doi.org/10.5281/zenodo.5244550.