CODEX System for Automated and Highly Multiplexed Immunofluorescence

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Project Summary Vanderbilt University Medical Center requests funds to purchase an Akoya CODEX (CO-Detection by indEXing) System that includes the CODEX fluidics instrument, CODEX processing computer, and a Keyence microscope with filters, to be placed in the Immunophenotyping Shared Resource (IPSR). Spatial resolution of the tissue microenvironment, particularly immune cell infiltration and their relationships with immune and stromal cells, is an essential approach for cancer biology, infectious disease, and autoimmune immunobiology inquiries, in both preclinical mechanistic studies and translational domains. The CODEX System allows for this comprehensive, spatially resolved, highly multiplexed biomarker analysis (40+ antigens) on formalin-fixed paraffin-embedded or frozen tissue. This highly multiplexed assay is achieved by conjugating barcoded oligonucleotides to antibodies followed by hybridization with a dye-labeled reporter for highly specific detection and efficient melting/rehybridization/fluorescent detection cycles. In 2019 the Vanderbilt Diabetes Research and Training Center and the Biomolecular Multimodal Imaging Center each purchased a CODEX System for their dedicated studies. Since acquisition, the systems are operating at capacity and lack the throughput to address the research of other Vanderbilt investigators. This lack of throughput severely limits the capability of cancer researchers and researchers studying non-diabetic autoimmune diseases to rapidly identify novel biomarkers or complex immunological states in tissues. The purchase of another CODEX System will greatly alleviate this research bottleneck. The CODEX System will support the ongoing NIH-funded research of 8 Major Users and 2 Minor Users across multiple departments at Vanderbilt University and Vanderbilt University Medical Center. These investigators have a critical need for high-dimensional in situ imaging analysis at cellular and subcellular levels for studying host- tumor interactions, immune cell subsets, and response and resistance to therapies. In addition to these investigator projects, we anticipate that many other NIH-funded projects will require the CODEX System to make ground-breaking discoveries, particularly those studying psoriasis, rheumatoid arthritis, lung inflammation (e.g. SARS-CoV-2 infection), and others where limited biopsies are possible. The expertise and support for the CODEX System at Vanderbilt are exceptional. The IPSR has a rich ten-year history of outstanding collaborative work studying host-tumor interactions, and has successful hands-on experience using the CODEX System on tumor tissue. The IPSR is co-led by three established faculty members with a wealth of experience in performing highly complex immunological assays, which is critical to the successful implementation of the CODEX System. Drs. Young Kim (Project PI) and Jeff Rathmell (Major User), IPSR Scientific Directors, and Dr. Kim Dahlman, IPSR Managing Director, along with experienced IPSR research team will provide sound guidance, assistance, and management of the CODEX System.