CODEX multiplex immunofluorescence imaging system

Rutgers Biomedical And Health Sciences, Newark NJ

Investigators

Abstract

Project Summary/Abstract: In this proposal we request funding to purchase an Akoya PhenoCycler Fusion imaging system for the Rutgers New Jersey Medical School (NJMS) Cellular Imaging and Histology Core facility. This system will provide the NJMS research community, as well as investigators in other Rutgers schools with state-of-the-art CO-Detection by IndEXing (CODEX) multiplex imaging technology. The Akoya PhenoCycler Fusion imaging system includes Akoya’s PhenoCycler, a custom designed automated microscope stage-top fluid exchange instrument and PhenoImager Fusion, an upright slide scanning epifluorescent microscope. This system will allow for the labeling of up to 60 individual biomarkers in single tissue section using oligonucleotide-conjugated antibodies against specific antigens. Tissues are stained once with all the barcoded primary antibodies, which can then be detected using complementary reporter DNA sequences conjugated to fluorescent dyes. Up to three of these fluorescent reporters can be imaged in a single CODEX cycle and then removed using a mild stripping buffer. Another CODEX cycle can then be started with three different oligonucleotides conjugated to the same reporters and the process can be repeated until all of the antigens of interest have been imaged. The investigators in this shared instrumentation grant have extensive experience using immunofluorescence or immunohistochemistry, and their research programs would substantially benefit from a multiplex imaging system to resolve spatial interactions between various cell types and immune cell subsets. Specifically, several investigators study the immune response to infection and inflammation, in which the recruitment or interactions of different innate and adaptive immune cells is involved in the pathology or resolution of disease. Additionally, several investigators are advancing our understanding of how specific cell types respond to local paracrine signals involved in antifungal immunity or within the nervous system. In all of these areas, the ability of the Akoya PhenoCycler Fusion imaging system to image up to 60 targets with single cell resolution will provide clear benefits over traditional fluorescence microscopy and provide the spatial information that is lacking from high-dimensional flow cytometry. Akoya’s ability to offer an integrated multiplex microscopic imaging system that minimizes damage to the tissue sections and is compatible with commercially available antibodies has led us to favor CODEX technology over other cyclic immunofluorescent techniques, such as Cytiva’s CellDive and Miltenyi’s MACSima platforms. Overall, the Akoya PhenoCycler Fusion imaging system would provide investigators at the New Jersey Medical School and the wider Rutgers community with much needed multiplex immunohistochemistry capabilities.