ArticleProceedings of the National Academy of Sciences of the United States of America2011
Targeting and imaging single biomolecules in living cells by complementation-activated light microscopy with split-fluorescent proteins.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 68 citations in OpenAlex.
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- Maximum likelihood-based estimation of diffusion coefficient is quick and reliable method for analyzing estradiol actions on surface receptor movements.Frontiers in neuroinformatics · 2023Article
- Emerin self-assembly and nucleoskeletal coupling regulate nuclear envelope mechanics against stress.Journal of cell science · 2022Article
- Cell-type-specific, multicolor labeling of endogenous proteins with split fluorescent protein tags inProceedings of the National Academy of Sciences of the United States of America · 2021Article
- TagBiFC technique allows long-term single-molecule tracking of protein-protein interactions in living cells.Communications biology · 2021Article
- Multiplexed labeling of cellular proteins with split fluorescent protein tags.Communications biology · 2021Article
- Split Green Fluorescent Protein-Based Contact Site Sensor (SPLICS ) for Heterotypic Organelle Juxtaposition as Applied to ER -Mitochondria Proximities.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Article
- GRP78 regulates CD44v membrane homeostasis and cell spreading in tamoxifen-resistant breast cancer.Life science alliance · 2019Article
- Development and Applications of Superfolder and Split Fluorescent Protein Detection Systems in Biology.International journal of molecular sciences · 2019Review
- Split Green Fluorescent Proteins: Scope, Limitations, and Outlook.Annual review of biophysics · 2019Review
- A fluorogenic array for temporally unlimited single-molecule tracking.Nature chemical biology · 2019Article
- LMO7 deficiency reveals the significance of the cuticular plate for hearing function.Nature communications · 2019Article
- Single-molecule imaging correlates decreasing nuclear volume with increasing TF-chromatin associations during zebrafish development.Nature communications · 2018Article
- Switchable Fluorophores for Single-Molecule Localization Microscopy.Chemical reviews · 2018Review
- Characterization of Split Fluorescent Protein Variants and Quantitative Analyses of Their Self-Assembly Process.Scientific reports · 2018Article
- Cellular imaging by targeted assembly of hot-spot SERS and photoacoustic nanoprobes using split-fluorescent protein scaffolds.Nature communications · 2018Article
- A split horseradish peroxidase for the detection of intercellular protein-protein interactions and sensitive visualization of synapses.Nature biotechnology · 2016Article
- Versatile protein tagging in cells with split fluorescent protein.Nature communications · 2016Article
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2 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Single-molecule (SM) microscopy allows outstanding insight into biomolecular mechanisms in cells. However, selective detection of single biomolecules in their native environment remains particularly challenging. Here, we introduce an easy methodology that combines specific targeting and nanometer accuracy imaging of individual biomolecules in living cells. In this method, named complementation-activated light microscopy (CALM), proteins are fused to dark split-fluorescent proteins (split-FPs), which are activated into bright FPs by complementation with synthetic peptides. Using CALM, the diffusion dynamics of a controlled subset of extracellular and intracellular proteins are imaged with nanometer precision, and SM tracking can additionally be performed with fluorophores and quantum dots. In cells, site-specific labeling of these probes is verified by coincidence SM detection with the complemented split-FP fusion proteins or intramolecular single-pair Förster resonance energy transfer. CALM is simple and combines advantages from genetically encoded and synthetic fluorescent probes to allow high-accuracy imaging of single biomolecules in living cells, independently of their expression level and at very high probe concentrations.
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