Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes

• DOI: 10.7554/elife.31657
• Publication Date: 2018-07-11T12:00:55Z
• Authors: Jia-Ren Lin; Benjamin Izar; Shu Wang; Clarence Yapp; Shaolin Mei; Parin M Shah; Sandro Santagata; Peter K Sorger
• Keywords: Immunofluorescence

The architecture of normal and diseased tissues strongly influences the development progression disease as well responsiveness resistance to therapy. We describe a tissue-based cyclic immunofluorescence (t-CyCIF) method for highly multiplexed immuno-fluorescence imaging formalin-fixed, paraffin-embedded (FFPE) specimens mounted on glass slides, most widely used histopathological diagnosis cancer other diseases. t-CyCIF generates up 60-plex images using an iterative process (a cycle) in which conventional low-plex fluorescence are repeatedly collected from same sample then assembled into high-dimensional representation. requires no specialized instruments or reagents is compatible with super-resolution imaging; we demonstrate its application quantifying signal transduction cascades, tumor antigens immune markers diverse tumors. simplicity adaptability makes it effective pre-clinical clinical research natural complement single-cell genomics.