Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.
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Abstract | 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. |
Year of Publication | 2018
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Journal | Elife
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Volume | 7
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Date Published | 2018 07 11
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ISSN | 2050-084X
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DOI | 10.7554/eLife.31657
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PubMed ID | 29993362
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PubMed Central ID | PMC6075866
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Grant list | R41 CA224503 / CA / NCI NIH HHS / United States
K08 CA222663 / CA / NCI NIH HHS / United States
P50 GM107618 / GM / NIGMS NIH HHS / United States
U54HL127365 / NH / NIH HHS / United States
K08CA222663 / NH / NIH HHS / United States
R41-CA224503 / NH / NIH HHS / United States
T32 GM008313 / GM / NIGMS NIH HHS / United States
P50GM107618 / NH / NIH HHS / United States
U54 HL127365 / HL / NHLBI NIH HHS / United States
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