ArticleBiophysical journal2010
Signal/noise analysis of FRET-based sensors.
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 26 papers.
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Who cites it
26 citing papers in PubMed, 46 citations in OpenAlex.
- In vivo quantitative FRET small animal imaging: Intensity versus lifetime-based FRET.Biophysical reports · 2023Article
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- Optimal inference of molecular interaction dynamics in FRET microscopy.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature Imaging with a Genetically Encoded Nanothermometer.Nano letters · 2022Article
- Measurement of neutral ceramidase activity in vitro and in vivo.Analytical biochemistry · 2022Review
- Native Cell Membrane Nanoparticles System for Membrane Protein-Protein Interaction Analysis.Journal of visualized experiments : JoVE · 2020Article
- Optically activated, customizable, excitable cells.PloS one · 2020Article
- Advanced FRET normalization allows quantitative analysis of protein interactions including stoichiometries and relative affinities in living cells.Scientific reports · 2019Article
- Live-Cell Imaging of Physiologically Relevant Metal Ions Using Genetically Encoded FRET-Based Probes.Cells · 2019Review
- Polymer Optical Fiber Tip Mass Production Etch Mechanism to Achieve CPC Shape for Improved Biosensor Performance.Sensors (Basel, Switzerland) · 2019Article
- Nitric Oxide Activates β-Cell Glucokinase by Promoting Formation of the "Glucose-Activated" State.Biochemistry · 2018Article
- SensorFRET: A Standardless Approach to Measuring Pixel-based Spectral Bleed-through and FRET Efficiency using Spectral Imaging.Scientific reports · 2017Article
- Optical sensors to gain mechanistic insights into signaling assemblies.Current opinion in structural biology · 2016Review
- Measurements of absolute concentrations of NADH in cells using the phasor FLIM method.Biomedical optics express · 2016Article
- New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories.Journal of biomolecular screening · 2016Article
- Two New FRET Imaging Measures: Linearly Proportional to and Highly Contrasting the Fraction of Active Molecules.PloS one · 2016Article
- MIiSR: Molecular Interactions in Super-Resolution Imaging Enables the Analysis of Protein Interactions, Dynamics and Formation of Multi-protein Structures.PLoS computational biology · 2015Article
- Polymer optical fiber compound parabolic concentrator tip for enhanced coupling efficiency for fluorescence based glucose sensors.Biomedical optics express · 2015Article
- Evaluating the performance of time-gated live-cell microscopy with lanthanide probes.Biophysical journal · 2015Article
- Plasmon-enhanced optical sensors: a review.The Analyst · 2015Review
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
Molecular sensors based on intramolecular Förster resonance energy transfer (FRET) have become versatile tools to monitor regulatory molecules in living tissue. However, their use is often compromised by low signal strength and excessive noise. We analyzed signal/noise (SNR) aspects of spectral FRET analysis methods, with the following conclusions: The most commonly used method (measurement of the emission ratio after a single short wavelength excitation) is optimal in terms of signal/noise, if only relative changes of this uncalibrated ratio are of interest. In the case that quantitative data on FRET efficiencies are required, these can be calculated from the emission ratio and some calibration parameters, but at reduced SNR. Lux-FRET, a recently described method for spectral analysis of FRET data, allows one to do so in three different ways, each based on a ratio of two out of three measured fluorescence signals (the donor and acceptor signal during a short-wavelength excitation and the acceptor signal during long wavelength excitation). Lux-FRET also allows for calculation of the total abundance of donor and acceptor fluorophores. The SNR for all these quantities is lower than that of the plain emission ratio due to unfavorable error propagation. However, if ligand concentration is calculated either from lux-FRET values or else, after its calibration, from the emission ratio, SNR for both analysis modes is very similar. Likewise, SNR values are similar, if the noise of these quantities is related to the expected dynamic range. We demonstrate these relationships based on data from an Epac-based cAMP sensor and discuss how the SNR changes with the FRET efficiency and the number of photons collected.
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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.