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

• Published in: eLife Vol. 7
• Main Authors: Lin, Jia-Ren, Izar, Benjamin, Wang, Shu, Yapp, Clarence, Mei, Shaolin, Shah, Parin M, Santagata, Sandro, Sorger, Peter K
• Format: Journal Article
• Language: English
• Published: England eLife Sciences Publications, Ltd 11.07.2018; eLife Sciences Publications Ltd
• Subjects: Antigens, Neoplasm - analysis; Cancer Biology; Computational and Systems Biology; Humans; Immunologic Factors - analysis; immunopathology; Microscopy, Fluorescence - methods; multiplexed imaging; Neoplasms - pathology; Signal Transduction; single-cell method; Tools and Resources; computational biology; systems biology; immunopathology; single-cell method; cancer biology; human; multiplexed imaging
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.31657

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.