Ultra High Content Analysis UHCA of Single Cells in Tissue 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 visualization Our goal is to create a suite
of integrated labeling display and analysis products to elucidate patterns of disease mechanisms in tissue which
is not possible using current methods To ensure this goal we have established a close partnership with Oregon
Health Science University OHSU and the commitment of a strong group of collaborators in the cancer research
community We have developed a novel cycIF technology based on antibody conjugated oligonucleotides Ab
oligos Briefly a single stranded oligo docking strand DS is conjugated to the primary antibody which can be
hybridized to a complementary single stranded oligo imaging strand IS conjugated to a conventional
fluorophore Each DS IS pair was designed to contain a restriction enzyme RE site for selective removal of
fluorescence permitting cycles of re staining and imaging Currently little functionality exists to optimally extract
the wealth of information in biomarker channel images We introduce a novel tool that can dynamically
visualize such images take advantage of the many channels for better cell segmentation even in complex tissue
and automatically correct and register the multiple cycles In Phase I a prototype of the imaging system will be
built and tested with a subset of labels Upon confirming feasibility Phase II will focus on the creation of specific
kits and software products in close harmony between biochemistry software and 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 laboratories Our goal is to create a suite of integrated labeling display
and analysis products to elucidate patterns of disease mechanisms in tissue which is not possible using current
methods which should help lead to more effectively strategies for therapy