An azide-based chemical cleavable linker has been introduced to design and synthesis novel fluorescent probes. These probes allow cyclic immunofluorescence staining which leads to the feasibility of highly multiplexed single cell in situ protein profiling. These highly multiplexed imaging-based platforms have the potential to quantify more than 100 protein targets in cultured cells and more than 50 protein targets in single cells in tissues.
This approach has been successfully applied in formalin-fixed paraffin-embedded (FFPE) brain tissues. Multiplexed protein expression level results reveal neuronal heterogeneity in the human hippocampus.
In 2004, a team of researchers at Tufts-New England
Medical Center in Boston, Massachusetts, investigated the fetal
cells that remained in the maternal blood stream after pregnancy.
The results were published in Transfer of Fetal Cells with
Multilineage Potential to Maternal Tissue. The team working on that
research included Kiarash Khosrotehrani, Kirby L. Johnson, Dong
Hyun Cha, Robert N. Salomon, and Diana W. Bianchi. The researchers
reported that the fetal cells passed to a pregnant woman during
pregnancy could develop into multiple cell types in her organs. They
studied these differentiated fetal cells in a cohort of women
fighting different diseases. The researchers found that the fetal
cells in the women differentiated into different cell types under
the influence of maternal tissues, and that those differentiated
cells concentrated in the tissue surrounding diseased tissues.
According to the team, this response could be a therapeutic response
to the disease in the once pregnant woman. The research indicated the long
lasting effects of pregnancy in a woman's body.
Novel approaches for highly multiplexed single cell in situ transcriptomic analysis were developed by our group to enable single-cell comprehensive RNA profiling in their native spatial contexts. Reiterative FISH was demonstrated to be able to detect >100 RNA species in single cell in situ, while more sophisticated approaches, consecutive FISH (C-FISH) and switchable fluorescent oligonucleotide based FISH (SFO-FISH), have the potential for whole transcriptome profiling at the single molecule sensitivity. The introduction of a cleavable fluorescent tyramide even enables sensitive RNA profiling in intact tissues with high throughput. These approaches will have wide applications in studies of systems biology, molecular diagnosis and targeted therapies.