OMAP-13: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Pancreas with CODEX, v1.0

OMAP-13 was designed for imaging of 4% PFA-fixed, cryopreserved human pancreas samples by CO-Detection by indEXing<br>(CODEX) (https://doi.org/10.1038/s41596-021-00556-8). Containing 27 antibodies and nuclear marker DAPI, this OMAP<br>provides spatial context for 13 anatomical structures and 21 cell types in the ASCT+B Pancreas table v1.2<br>(https://doi.org/10.48539/HBM975.BBTF.393) and shares 8 markers with OMAP-6<br>(https://doi.org/10.48539/HBM868.XLTM.922). It also facilitates identification of gamma and epsilon cells, lymphatic<br>endothelial cells, pericytes, resident immune cells, neuronal structures, and some stromal cells and the extracellular<br>matrix (ECM). Due to the inclusion of biomarkers PPY, GHRL, PTPRC (CD45), CD19, ITGAX (CD11c), AIF1 (IBA1), and CD14,<br>this panel is suitable for all regions of the human pancreas (including the uncinate process, UBERON:0010373) from<br>birth to adulthood and can detect inflammatory cells present in diabetes. Panel design and validation was performed on<br>the PhenoCycler Open and Fusion systems (Akoya Biosciences). Carrier free aliquots for antibodies directed against<br>targets GHRL, PPY, and LYVE1 were ordered directly from R&D Biosciences to allow custom conjugation; antibody<br>metadata reflect the catalog versions. The GP2 antibody, which was purified and conjugated by Leinco Technologies,<br>Inc., is best used in conjunction with another acinar cell marker. Application of this CODEX panel to early-stage T2D<br>samples (Walker et al. 2021; https://doi.org/10.1101/2021.12.16.466282) included cellular segmentation by DAPI using<br>the Highplex FL module of HALO image analysis software (Indica Labs). Details on sample preparation and antibody<br>screening can be found at https://doi.org/10.17504/protocols.io.36wgq7dryvk5/v1 and a representative dataset is<br>uploaded at https://doi.org/10.5281/zenodo.7916784. Interactive image browsing is available on Pancreatlas<br>(RRID:SCR_01856).