ArticlePloS one2009
Simultaneous live cell imaging using dual FRET sensors with a single excitation light.
Article in PloS one, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.
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Who cites it
42 citing papers in PubMed, 115 citations in OpenAlex.
- High-throughput screening of FRET-based proteins using a hyperspectral microcapillary array.iScience · 2026Article
- Live imaging of apoptotic signaling flow using tunable combinatorial FRET-based bioprobes for cell population analysis of caspase cascades.Scientific reports · 2022Article
- An ultra-small nine-color spectrometer with a two-layer biparted ten-dichroic-mirror array and an image sensor.Scientific reports · 2022Article
- CNP regulates cardiac contractility and increases cGMP near both SERCA and TnI: difference from BNP visualized by targeted cGMP biosensors.Cardiovascular research · 2022Article
- A Real-Time, Plate-Based BRET Assay for Detection of cGMP in Primary Cells.International journal of molecular sciences · 2022Article
- CRISPR/Cas9-mediated generation and analysis of N terminus polymorphic models of βMolecular therapy. Methods & clinical development · 2021Article
- CUTie2: The Attack of the Cyclic Nucleotide Sensor Clones.Frontiers in molecular biosciences · 2021Article
- Mind the gap (junction): cGMP induced by nitric oxide in cardiac myocytes originates from cardiac fibroblasts.British journal of pharmacology · 2019Article
- Dual-FRET imaging of IPScientific reports · 2019Article
- FRET-based cyclic GMP biosensors measure low cGMP concentrations in cardiomyocytes and neurons.Communications biology · 2019Article
- Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy.Redox biology · 2018Article
- Distinct submembrane localisation compartmentalises cardiac NPR1 and NPR2 signalling to cGMP.Nature communications · 2018Article
- cGMP Signaling in the Cardiovascular System-The Role of Compartmentation and Its Live Cell Imaging.International journal of molecular sciences · 2018Review
- Imaging of PDE2- and PDE3-Mediated cGMP-to-cAMP Cross-Talk in Cardiomyocytes.Journal of cardiovascular development and disease · 2018Review
- Cardiac optogenetics: using light to monitor cardiac physiology.Basic research in cardiology · 2017Review
- Compartmentalized cGMP Responses of Olfactory Sensory Neurons inThe Journal of neuroscience : the official journal of the Society for Neuroscience · 2017Article
- FRET from single to multiplexed signaling events.Biophysical reviews · 2017Review
- Multiplexing PKA and ERK1&2 kinases FRET biosensors in living cells using single excitation wavelength dual colour FLIM.Scientific reports · 2017Article
- A Guide to Fluorescent Protein FRET Pairs.Sensors (Basel, Switzerland) · 2016Review
- Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells.International journal of molecular sciences · 2015Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescence resonance energy transfer (FRET) between fluorescent proteins is a powerful tool for visualization of signal transduction in living cells, and recently, some strategies for imaging of dual FRET pairs in a single cell have been reported. However, these necessitate alteration of excitation light between two different wavelengths to avoid the spectral overlap, resulting in sequential detection with a lag time. Thus, to follow fast signal dynamics or signal changes in highly motile cells, a single-excitation dual-FRET method should be required. Here we reported this by using four-color imaging with a single excitation light and subsequent linear unmixing to distinguish fluorescent proteins. We constructed new FRET sensors with Sapphire/RFP to combine with CFP/YFP, and accomplished simultaneous imaging of cAMP and cGMP in single cells. We confirmed that signal amplitude of our dual FRET measurement is comparable to of conventional single FRET measurement. Finally, we demonstrated to monitor both intracellular Ca(2+) and cAMP in highly motile cardiac myocytes. To cancel out artifacts caused by the movement of the cell, this method expands the applicability of the combined use of dual FRET sensors for cell samples with high motility.
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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.