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A Simple Open-source Method For Highly Multiplexed Imaging Of Single Cells In Tissues And Tumours

Jia-Ren Lin, Benjamin Izar, Shaolin Mei, S. Wang, Parin Shah, P. Sorger
Published 2017 · Biology

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Intratumoural heterogeneity strongly influences the development and progression of cancer as well as responsiveness and resistance to therapy. To improve our ability to measure and analyze such heterogeneity we have developed an open source method for fluorescence imaging of up to 60 protein antigens at subcellular resolution using formalin-fixed, paraffin-embedded (FFPE) tissue samples mounted on glass slides, the most widely used specimens for the diagnosis of cancer and other diseases. As described here, tissue-based cyclic immunofluorescence (t-CyCIF) creates high-dimensional imaging data through successive acquisition of four-color images and requires no specialized instruments or reagents. We apply t-CyCIF to 14 cancer and healthy tissue types and quantify the extent of cell to cell variability in signal transduction cascades, tumor antigens and stromal markers. By imaging immune cell lineage markers we enumerate classes of tumour-infiltrating lymphocytes (TILs) and their spatial relationships to the tumor microenvironment (TME). The simplicity and adaptability of t-CyCIF makes it a powerful method for pre-clinical and clinical research and a natural complement to single-cell genomics.
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