ArticleBiophysical journal2010
Three-color spectral FRET microscopy localizes three interacting proteins in living cells.
Article in Biophysical journal, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 66 citations in OpenAlex.
- Development of three-colour FRET cascade for force sensing of the putative RIAM-vinculin interaction in fibroblasts.Communications chemistry · 2025Article
- Genetically Encoded Fluorescence Barcodes Allow for Single-Cell Analysis via Spectral Flow Cytometry.ACS synthetic biology · 2025Article
- Single-photon smFRET. I: Theory and conceptual basis.Biophysical reports · 2023Article
- Three-Fluorophore FRET Enables the Analysis of Ternary Protein Association in Living Plant Cells.Plants (Basel, Switzerland) · 2022Article
- Flow cytometry based-FRET: basics, novel developments and future perspectives.Cellular and molecular life sciences : CMLS · 2022Review
- Multicolor fluorescence fluctuation spectroscopy in living cells via spectral detection.eLife · 2021Article
- A Combined Acceptor Photobleaching and Donor Fluorescence Lifetime Imaging Microscopy Approach to Analyze Multi-Protein Interactions in Living Cells.Frontiers in molecular biosciences · 2021Article
- Of numbers and movement - understanding transcription factor pathogenesis by advanced microscopy.Disease models & mechanisms · 2020Review
- Application of FRET Biosensors in Mechanobiology and Mechanopharmacological Screening.Frontiers in bioengineering and biotechnology · 2020Review
- Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity.eLife · 2019Article
- Monitoring ligand-dependent assembly of receptor ternary complexes in live cells by BRETFect.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Multicolor protein FRET with tryptophan, selective coumarin-cysteine labeling, and genetic acridonylalanine encoding.Chemical communications (Cambridge, England) · 2017Article
- FRET from single to multiplexed signaling events.Biophysical reviews · 2017Review
- Optical sensors to gain mechanistic insights into signaling assemblies.Current opinion in structural biology · 2016Review
- A Guide to Fluorescent Protein FRET Pairs.Sensors (Basel, Switzerland) · 2016Review
- Techniques for the Analysis of Protein-Protein Interactions in Vivo.Plant physiology · 2016Review
- Multiplexed 3D FRET imaging in deep tissue of live embryos.Scientific reports · 2015Article
- Optimizing fluorescent protein trios for 3-Way FRET imaging of protein interactions in living cells.Scientific reports · 2015Article
- Three-color confocal Förster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2015Article
- Application of fluorescence resonance energy transfer in protein studies.Journal of molecular structure · 2014Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
FRET technologies are now routinely used to establish the spatial relationships between two cellular components (A and B). Adding a third target component (C) increases the complexity of the analysis between interactions AB/BC/AC. Here, we describe a novel method for analyzing a three-color (ABC) FRET system called three-color spectral FRET (3sFRET) microscopy, which is fully corrected for spectral bleedthrough. The approach quantifies FRET signals and calculates the apparent energy transfer efficiencies (Es). The method was validated by measurement of a genetic (FRET standard) construct consisting of three different fluorescent proteins (FPs), mTFP, mVenus, and tdTomato, linked sequentially to one another. In addition, three 2-FP reference constructs, tethered in the same way as the 3-FP construct, were used to characterize the energy transfer pathways. Fluorescence lifetime measurements were employed to compare the relative relationships between the FPs in cells producing the 3-FP and 2-FP fusion proteins. The 3sFRET microscopy method was then applied to study the interactions of the dimeric transcription factor C/EBPalpha (expressing mTFP or mVenus) with the heterochromatin protein 1alpha (HP1alpha, expressing tdTomato) in live-mouse pituitary cells. We show how the 3sFRET microscopy method represents a promising live-cell imaging technique to monitor the interactions between three labeled cellular components.
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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.