OMAP-23: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of vermiform appendix (FFPE) with MICS on MACSima

Description

 OMAP-23 was designed for MICS (MACSima imaging cyclic staining) imaging of FFPE human vermiform appendix sample.  Tissue fixation and antigen retrieval is described in (Spatial protein and RNA analysis on the same tissue section using MICS technology). The MACSima technology is described in detail in the following publication (MACSima imaging cyclic staining (MICS) technology reveals combinatorial target pairs for CAR T cell treatment of solid tumors). All, antibodies in this panel are recombinant antibodies with a mutated human IgG1 constant region, removing Fc receptor binding capacity of human IgG1, eliminating the need for additional blocking steps and reducing non-specific binding. The use of human IgG1 recombinant antibodies allows for the addition of uncoupled monoclonal antibodies from other species followed by a fluorescence labelled secondary reagent specific for the species of the monoclonal antibody. The multiplex system has been described already for OMAP-10 and OMAP-21. A new dye VioB515 is used for one reagents. The fluorescence is removed by cleavage. The panel contains 28 antibodies and the nuclear marker DAPI for image alignment and nuclear segmentation. This OMAP provides a spatial context for six anatomical structures and most cell types present in the vermiform appendix (link to ASCT+B table added). OMAP-23 follows OMAP-21, with fewer antibodies but adding antibodies for non-immune cells.

All reagents are from Miltenyi Biotec and have been rigorously tested through an internal quality control system to have minimal variation between lots. For this reason, lot information is not included in this table. Analysis was performed by an accompanied software package MACSIQ View Analysis also described in the MACSima publication (https://doi.org/10.1038/s41598-022-05841-4). The MACSima system is continuously evolving, this is the third OMAP for the MACSima system. A representative dataset created using OMAP-23 can be found here 10.5281/zenodo.14008816 .The AVRs for the dataset can be found here (to be added).