ArticleMolecules (Basel, Switzerland)2021
Multiplexed In Situ Protein Profiling with High-Performance Cleavable Fluorescent Tyramide.
Article in Molecules (Basel, Switzerland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Emerging technologies in the spatial proteomics landscape.Analytical and bioanalytical chemistry · 2026Review
- TyrRu: A Tyramide-Amplified Multimodal Ruthenium Tag for Elemental Mass Spectrometry and Immunofluorescence Imaging.Chemical & biomedical imaging · 2026Article
- Highly Sensitive Spatial Proteomics with Multicolor Cleavable Fluorescent Tyramide.Chemical & biomedical imaging · 2025Article
- Single-cell spatial proteomics.Histology and histopathology · 2025Review
- Ultrasensitive and Multiplexed Protein Imaging with Clickable and Cleavable Fluorophores.Analytical chemistry · 2024Article
- Ultrasensitive and multiplexed protein imaging with clickable and cleavable fluorophores.bioRxiv : the preprint server for biology · 2023Article
- Multiplex Protein Imaging through PACIFIC: Photoactive Immunofluorescence with Iterative Cleavage.ACS bio & med chem Au · 2023Article
- Multiplexed Single-Cell in Situ Protein Profiling.ACS measurement science au · 2022Review
- Simultaneous amplification of multiple immunofluorescence signals via cyclic staining of target molecules using mutually cross-adsorbed antibodies.Scientific reports · 2022Article
- Ultrasensitive and Multiplexed Protein Imaging with Cleavable Fluorescent Tyramide and Antibody Stripping.International journal of molecular sciences · 2021Article
- Highly Sensitive and Multiplexed In Situ RNA Profiling with Cleavable Fluorescent Tyramide.Cells · 2021Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Understanding the composition, function and regulation of complex cellular systems requires tools that quantify the expression of multiple proteins at their native cellular context. Here, we report a highly sensitive and accurate protein in situ profiling approach using off-the-shelf antibodies and cleavable fluorescent tyramide (CFT). In each cycle of this method, protein targets are stained with horseradish peroxidase (HRP) conjugated antibodies and CFT. Subsequently, the fluorophores are efficiently cleaved by mild chemical reagents, which simultaneously deactivate HRP. Through reiterative cycles of protein staining, fluorescence imaging, fluorophore cleavage, and HRP deactivation, multiplexed protein quantification in single cells in situ can be achieved. We designed and synthesized the high-performance CFT, and demonstrated that over 95% of the staining signals can be erased by mild chemical reagents while preserving the integrity of the epitopes on protein targets. Applying this method, we explored the protein expression heterogeneity and correlation in a group of genetically identical cells. With the high signal removal efficiency, this approach also enables us to accurately profile proteins in formalin-fixed paraffin-embedded (FFPE) tissues in the order of low to high and also high to low expression levels.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.