OMAP-14: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Kidney with CyCIF, v1.0

OMAP-14 was designed for CyCIF imaging of unfixed frozen human kidney samples<br>(https://dx.doi.org/10.17504/protocols.io.81wgb1m3yvpk/v3). The panel contains 17 antibodies, 1 lectin, and Hoechst<br>33342 for image alignment and nuclear segmentation. The OMAP provides a spatial context for the main anatomical<br>structures and key cell types present in the healthy human kidney. These include markers for podocytes, mesangial and<br>endothelial cells, glomerular basement membrane, mesangial matrix in glomeruli, endothelial and vascular smooth<br>muscle cells in the renal vasculature, renal tubular segments including proximal tubular epithelial cells, loops of<br>Henle (including descending thin limb and thick ascending limb), distal convoluted tubules, connecting segment,<br>cortical and medullary collecting duct principal cells and intercalated cells, myofibroblasts and peritubular<br>extracellular matrix. A polyclonal antibody recognizing Zonula occludens-1 ZO-1, also known as Tight junction<br>protein-1, antibodies, which detect tight junctions, were used to segment podocyte plasma membranes as well as<br>basolateral plasma membranes in all tubular epithelium distal to proximal tubules. Lotus Tetragonolobus Lectin (LTL)<br>was used to visualize proximal tubular epithelial cells, collecting duct intercalated cells, and inner medullary<br>collecting duct principal cells. LTL is visualized in the accompanying datasets but not included in the OMAP because<br>it is not an antibody. Additional details on sample preparation (https://dx.doi.org/10.17504/protocols.io.bfskjncw)<br>and custom conjugation of antibodies (https://dx.doi.org/10.17504/protocols.io.667hhhn) are included in these links.<br>Representative datasets including two tubular segment and one glomerular segmentation 2D CyCIF images, and one<br>glomerular segmentation 3D image have been uploaded here:https://dx.doi.org/10.5281/zenodo.10407946;<br>https://dx.doi.org/10.5281/zenodo.10440320;<br>https://dx.doi.org/10.5281/zenodo.10445538; https://dx.doi.org/10.5281/zenodo.10445538;<br>https://dx.doi.org/10.5281/zenodo.10447488 ; https://dx.doi.org/10.5281/zenodo.10445551.