OMAP-1 Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Lymph Node with IBEX, v1.2

OMAP-1 was designed for IBEX (Iterative Bleaching Extends multi-pleXity) imaging of fixed frozen human lymph nodes<br>(https://doi.org/10.1038/s41596-021-00644-9; https://doi.org/10.1073/pnas.2018488117). The panel contains 39<br>antibodies and the nuclear marker Hoechst 33342 for image alignment and nuclear segmentation. his OMAP provides a<br>spatial context for all anatomical structures and most cell types present in the [ASCT+B lymph node table,<br>v1.1](https://doi.org/10.48539/HBM573.SHCQ.259). Additionally, the inclusion of protein biomarkers BCL2, CD10, CD138,<br>and CD44 allow profiling of lymph node disease states, e.g. follicular lymphoma and metastasis. The core and<br>prioritized OMAP targets detailed here overlap (~50-85%) with panels developed for multiplexed imaging of lymphoid<br>samples using other technologies and sample preparations (https://doi.org/10.7554/eLife.31657,<br>https://doi.org/10.1002/eji.202048891). This OMAP does not contain antibodies for typing pericytes, plasmacytoid<br>dendritic cells, or different T helper lineages (Th1, Th2, Th17). Beyond spatial mapping of healthy human lymph nodes,<br>this OMAP was used to characterize the tissue microenvironment of follicular lymphoma lymph nodes. For this study,<br>object-based cellular segmentation was performed using a convolutional neural network (CNN)-based approach with Mask<br>R-CNN architecture and ResNet-50 as a backbone. As an input three channels were used: Hoechst for nuclei, CD45 as a<br>base membrane, and composite of several other membrane markers (CD138, CD163, CD94, CD69, CD8, CD4). More details and<br>accompanying datasets associated with OMAP-1 can be found here (https://doi.org/10.1101/2022.06.03.494716,<br>10.5281/zenodo.6629388). Several antibodies required custom conjugation by commercial suppliers or antibody<br>conjugation kits (e.g., AF532, Thermo A20182).