Ultra-High Content Analysis (UHCA) of Single Cells in Tissue: 60+ channel immunofluorescence labeling kits and companion imaging software for everyone

We propose to create a modular suite of products that will make highly multiplexed imaging of individual cells in intact tissue available to a wide range of laboratories through validated labeling kits and companion imaging software that promotes discovery through interactive image and data visualizationOur goal is to create a suite of integrated labelingdisplay and analysis products to elucidate patterns of disease mechanisms in tissuewhich is not possible using current methodsTo ensure this goalwe have established a close partnership with Oregon Health Science UniversityOHSUand the commitment of a strong group of collaborators in the cancer research communityWe have developed a novel cycIF technology based on antibody conjugated oligonucleotidesAboligosBrieflya single stranded oligodocking strandDSis conjugated to the primary antibody which can be hybridized to a complementary single stranded oligoimaging strandISconjugated to a conventional fluorophoreEach DS IS pair was designed to contain a restriction enzymeREsite for selective removal of fluorescence permitting cycles of re staining and imagingCurrentlylittle functionality exists to optimally extract the wealth of information inbiomarker channel imagesWe introduce a novel tool that can dynamically visualize such imagestake advantage of the many channels for better cell segmentation even in complex tissueand automatically correct and register the multiple cyclesIn Phase Ia prototype of the imaging system will be built and tested with a subset of labelsUpon confirming feasibilityPhase II will focus on the creation of specific kits and software products in close harmony between biochemistrysoftwareand research needs We propose to create a modular suite of products that will make highly multiplexed imaging of individual cells in intact tissue available to a wide range of laboratoriesOur goal is to create a suite of integrated labelingdisplay and analysis products to elucidate patterns of disease mechanisms in tissuewhich is not possible using current methodswhich should help lead to more effectively strategies for therapy