Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.
Jia-Ren LinBenjamin IzarShu WangClarence YappShaolin MeiParin M ShahSandro SantagataPeter Karl SorgerPublished in: eLife (2018)
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.
Keyphrases
- single cell
- fluorescence imaging
- high resolution
- rna seq
- gene expression
- deep learning
- photodynamic therapy
- convolutional neural network
- high throughput
- optical coherence tomography
- papillary thyroid
- palliative care
- dendritic cells
- stem cells
- induced pluripotent stem cells
- machine learning
- fine needle aspiration
- image quality
- magnetic resonance
- pluripotent stem cells
- high speed
- quantum dots