ArticleScientific reports2017
SensorFRET: A Standardless Approach to Measuring Pixel-based Spectral Bleed-through and FRET Efficiency using Spectral Imaging.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Detection of fluorescent protein mechanical switching in cellulo.Cell reports methods · 2024Article
- Genetically Encoded Fluorescent Probe for Detection of Heme-Induced Conformational Changes in Cytochrome c.Biosensors · 2023Article
- Nuclear lamina strain states revealed by intermolecular force biosensor.Nature communications · 2023Article
- Visualizing Neurons Under Tension In Vivo with Optogenetic Molecular Force Sensors.Methods in molecular biology (Clifton, N.J.) · 2023Article
- The LINC complex is required for endothelial cell adhesion and adaptation to shear stress and cyclic stretch.Molecular biology of the cell · 2021Article
- Imaging methods in mechanosensing: a historical perspective and visions for the future.Molecular biology of the cell · 2021Review
- A hybrid model of intercellular tension and cell-matrix mechanical interactions in a multicellular geometry.Biomechanics and modeling in mechanobiology · 2020Article
- Interclass GPCR heteromerization affects localization and trafficking.Science signaling · 2020Article
- Fly-QMA: Automated analysis of mosaic imaginal discs in Drosophila.PLoS computational biology · 2020Article
- Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells.Cellular and molecular bioengineering · 2019Article
- Article
- Using Nesprin Tension Sensors to Measure Force on the LINC Complex.Methods in molecular biology (Clifton, N.J.) · 2018Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Fluorescence microscopy of FRET-based biosensors allow nanoscale interactions to be probed in living cells. This paper describes a novel approach to spectrally resolved fluorescence microscopy, termed sensorFRET, that enables quantitative measurement of FRET efficiency. This approach is an improvement on existing methods (FLIM, sRET, luxFRET, pFRET), as it does not require single fluorophore standards to be measured with every experiment and the acquisition is intensity independent, allowing the laser power to be optimized for varying levels of fluorophore expression. Additionally, it was found that all spectral based methods, including sensorFRET, fail at specific fluorophore-excitation wavelength combinations. These combinations can be determined a priori using sensorFRET, whereas other methods would give no indication of inaccuracies. This method was thoroughly validated and compared to existing methods using simulated spectra, Fluorescein and TAMRA dye mixtures as a zero FRET control, and Cerulean-Venus FRET standards as positive FRET controls. Simulations also provided a means of quantifying the uncertainty in each measurement by relating the fit residual of noisy spectra to the standard deviation of the measured FRET efficiency. As an example application, Teal-Venus force sensitive biosensors integrated into E-cadherin were used to resolve piconewton scale forces along different parts of an individual cell junction.
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