OMAP-9: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Kidney with CODEX, v1.2

OMAP-9 was designed for CODEX multiplex antibody-based imaging of fresh frozen human kidney tissue<br>(dx.doi.org/10.17504/protocols.io.8epv5jdbnl1b/v1). The panel contains 41 antibodies and the nuclear marker DAPI for<br>image alignment and nuclear segmentation. OMAP-9 provides a spatial context for all anatomical structures and most<br>cell types present in the ASCT+B kidney table v1.1. This OMAP expands on OMAP-3 (10.48539/HBM373.TBKP.375), an 18-plex<br>kidney CODEX panel, by including additional markers for granular phenotyping of key cells in the kidney such as<br>structural cells (Podocalyxin, LRP2, E-cadherin, SLC12A3, Na/K ATPase and Cytokeratin 8), immune cells including T<br>cells (CD3D, CD4, CD45RO, CD8, FOXP3, GATA-3), myeloid cells (CD11c, CD68, CD206, HLA-DR, Myeloperoxidase), and B<br>cells (CD20). Antibodies directed against injury markers (KIM1, IGFBP7, Osteopontin, PLSCR1, VCAM1, MCP1, phosphor<br>c-JUN, phosphor MLKL, Fibronectin, Collagen IV, LC3, Cystatin 3, and PROM1) allows profiling of kidney disease<br>progression, prognosis, and repair. The following markers utilize different antibody clones than OMAP-3: vimentin,<br>beta-catenin, and uromodulin. Mesangial cells present in the glomerulus are not targeted in the existing panel due to<br>the absence of suitable antibodies compatible with the methodology. Additional details on sample preparation and CODEX<br>multiplexing workflow can be found on Protocols.io (dx.doi.org/10.17504/protocols.io.8epv5jdbnl1b/v1). Correction to<br>update to authors list and spelling of names appears in OMAP-9 v1.1.OMAP-9 专为新鲜冰冻人肾脏组织的CODEX多重抗体成像而设计（相关实验方案详见dx.doi.org/10.17504/protocols.io.8epv5jdbnl1b/v1）。该标记组合包含41种抗体以及用于图像配准与细胞核分割的核标记物DAPI。OMAP-9可为ASCT+B肾脏表格v1.1中收录的所有解剖结构与多数细胞类型提供空间背景信息。 相较于此前的18重肾脏CODEX标记组合OMAP-3（编号：10.48539/HBM373.TBKP.375），本OMAP-9新增了一系列标记物，可对肾脏关键细胞进行精细化表型分型：包括结构细胞（Podocalyxin、LRP2、E-钙粘蛋白、SLC12A3、Na/K ATP酶以及细胞角蛋白8）、免疫细胞（其中T细胞相关标记包括CD3D、CD4、CD45RO、CD8、FOXP3、GATA-3；髓系细胞相关标记包括CD11c、CD68、CD206、HLA-DR、髓过氧化物酶；B细胞相关标记包括CD20）。 针对损伤标记物（KIM1、IGFBP7、骨桥蛋白、PLSCR1、VCAM1、MCP1、磷酸化c-JUN、磷酸化MLKL、纤连蛋白、IV型胶原、LC3、胱抑素3以及PROM1）的抗体，可用于分析肾脏疾病的进展、预后与修复过程。 波形蛋白、β-连环蛋白以及尿调蛋白这三种标记物所使用的抗体克隆与OMAP-3存在差异。 由于缺乏适配该实验方法的合适抗体，当前标记组合未靶向肾小球内的系膜细胞。 关于样本制备与CODEX多重成像流程的更多细节，可参阅Protocols.io平台上的相关方案（dx.doi.org/10.17504/protocols.io.8epv5jdbnl1b/v1）。 OMAP-9 v1.1版本已针对作者列表及姓名拼写错误进行了修正。