ArticleNature biotechnology2022
Spatiotemporal multiplexed immunofluorescence imaging of living cells and tissues with bioorthogonal cycling of fluorescent probes.
Article in Nature biotechnology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed.
- Achieving cell-type-specific bioorthogonal chemistry using enzyme-activated caged tetrazines.Nature chemical biology · 2026Article
- Optically switchable CLEAR probes enable rapid, biocompatible and high-efficiency fluorophore exchange for ultra-plex, high-resolution immunofluorescence imaging.Chemical science · 2026Article
- Single-Cell Protein Assays in Context: From 2D to 3D and In Situ Analysis.Annual review of analytical chemistry (Palo Alto, Calif.) · 2026Review
- Article
- Dynamic Decoration of DNA Scaffolds for High-Resolution Cancer Cell Subtyping.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Single-molecule validation and optimised protocols for the use of secondary nanobodies in multiplexed immunoassays.Journal of microscopy · 2026Article
- Spatiotemporally controlled drug release via a click-release system utilizing mono-alkyl-hydroxylamine and cyclooctyne chemistry.Nature communications · 2026Article
- Positioning and reversible suppression of CCR7Immunity · 2026Article
- Unified platform for multiplex immunofluorescence across liver tissues and engineered models.eGastroenterology · 2026Article
- Integrative Biosensing Nanoplasmonic Array for Real-Time Spatiotemporal Imaging of Protein Secretion in Cell-to-Cell Communication.Small science · 2025Article
- Targeting cancer-associated fibroblasts for real-time intraoperative tumor identification with a spray-on fluorescent probe.Science advances · 2025Article
- Scalable topic modelling decodes spatial tissue architecture for large-scale multiplexed imaging analysis.Nature communications · 2025Article
- Temporal and spatial omics technologies for 4D profiling.Nature methods · 2025Review
- Patient-derived organotypic tumor spheroids, tumoroids, and organoids: advancing immunotherapy using state-of-the-art 3D tumor model systems.Lab on a chip · 2025Review
- Imaging 3D cell cultures with optical microscopy.Nature methods · 2025Review
- Spatial analysis identifies DC niches as predictors of pembrolizumab therapy in head and neck squamous cell cancer.Cell reports. Medicine · 2025Article
- Large-scale combinatorial optical barcoding of cells with laser particles.Light, science & applications · 2025Article
- AI-based hardware and software tools in microscopy to boost research in immunology and virology.Frontiers in immunology · 2025Review
- Advances and challenges in precision imaging.The Lancet. Oncology · 2025Review
- Synthesis of CThe Journal of organic chemistry · 2024Article
Corrections and comments
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Authors and funding
11 authors.
Funding
Abstract
Cells in complex organisms undergo frequent functional changes, but few methods allow comprehensive longitudinal profiling of living cells. Here we introduce scission-accelerated fluorophore exchange (SAFE), a method for multiplexed temporospatial imaging of living cells with immunofluorescence. SAFE uses a rapid bioorthogonal click chemistry to remove immunofluorescent signals from the surface of labeled cells, cycling the nanomolar-concentration reagents in seconds and enabling multiple rounds of staining of the same samples. It is non-toxic and functional in both dispersed cells and intact living tissues. We demonstrate multiparameter (n ≥ 14), non-disruptive imaging of murine peripheral blood mononuclear and bone marrow cells to profile cellular differentiation. We also show longitudinal multiplexed imaging of bone marrow progenitor cells as they develop into neutrophils over 6 days and real-time multiplexed cycling of living mouse hepatic tissues. We anticipate that SAFE will find broad utility for investigating physiologic dynamics in living systems.
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Registered trials
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