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

Elife
Authors
Keywords
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
Journal
Elife
Volume
7
Date Published
2018 07 11
ISSN
2050-084X
DOI
10.7554/eLife.31657
PubMed ID
29993362
PubMed Central ID
PMC6075866
Links
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