2018/07/11 by Jia‐Ren Lin, Benjamin Izar, Shu Wang +5 · 4 citations
Biochemistry, Genetics and Molecular Biology · Immunology and Microbiology · #Advanced Fluorescence Microscopy Techniques #Immunotherapy and Immune Responses #Single-cell and spatial transcriptomics
paper · doi:10.7554/elife.31657
openalex publication_date 2018/07/11 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/03
The architecture of normal and diseased tissues strongly influences the development and progression of disease as well as responsiveness and resistance to therapy. We describe a tissue-based cyclic immunofluorescence (t-CyCIF) method for highly multiplexed immuno-fluorescence imaging of formalin-fixed, paraffin-embedded (FFPE) specimens mounted on glass slides, the most widely used specimens for histopathological diagnosis of cancer and other diseases. t-CyCIF generates up to 60-plex images using an iterative process (a cycle) in which conventional low-plex fluorescence images are repeatedly collected from the same sample and then assembled into a high-dimensional representation. t-CyCIF requires no specialized instruments or reagents and is compatible with super-resolution imaging; we demonstrate its application to quantifying signal transduction cascades, tumor antigens and immune markers in diverse tissues and tumors. The simplicity and adaptability of t-CyCIF makes it an effective method for pre-clinical and clinical research and a natural complement to single-cell genomics.