ArticleBiophysical journal2008
Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy.
Article in Biophysical journal, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Phasor and neural network approaches for rapid fluorophore fraction analysis in temporal-spectral multiplexed data.Journal of biomedical optics · 2025Article
- Multiplexed In Vivo Imaging with Fluorescence Lifetime-Modulating Tags.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Artificial Neural Network (ANN)-Based Determination of Fractional Contributions from Mixed Fluorophores using Fluorescence Lifetime Measurements.Journal of fluorescence · 2024Article
- Lifetime-based analysis of binary fluorophores mixtures in the low photon count limit.iScience · 2022Article
- Separation of spectrally overlapping fluorophores using intra-exposure excitation modulation.Biophysical reports · 2021Review
- A Guide to Fluorescence Lifetime Microscopy and Förster's Resonance Energy Transfer in Neuroscience.Current protocols in neuroscience · 2020Review
- Hyperspectral multiphoton microscopy forOptica · 2020Article
- UNMIX-ME: spectral and lifetime fluorescence unmixing via deep learning.Biomedical optics express · 2020Article
- Bright GFP with subnanosecond fluorescence lifetime.Scientific reports · 2018Article
- Quadratic blind linear unmixing: A graphical user interface for tissue characterization.Computer methods and programs in biomedicine · 2016Article
- Green fluorescent protein with anionic tryptophan-based chromophore and long fluorescence lifetime.Biophysical journal · 2015Article
- Digitally synthesized beat frequency-multiplexed fluorescence lifetime spectroscopy.Biomedical optics express · 2014Article
- Reversible switching between nonquenched and quenched states in nanoscale linear arrays of plant light-harvesting antenna complexes.Langmuir : the ACS journal of surfaces and colloids · 2014Article
- Estimation of the number of fluorescent end-members for quantitative analysis of multispectral FLIM data.Optics express · 2014Article
- FRET-FLIM applications in plant systems.Protoplasma · 2014Review
- Article
- Application of non-negative matrix factorization to multispectral FLIM data analysis.Biomedical optics express · 2012Article
- Imaging the coordination of multiple signalling activities in living cells.Nature reviews. Molecular cell biology · 2011Review
- Phase differential enhancement of FLIM to distinguish FRET components of a biosensor for monitoring molecular activity of Membrane Type 1 Matrix Metalloproteinase in live cells.Journal of fluorescence · 2011Article
- Non fitting based FRET-FLIM analysis approaches applied to quantify protein-protein interactions in live cells.Biophysical reviews · 2011Review
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4 authors.
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Abstract
Fluorescence lifetime imaging microscopy (FLIM) is a quantitative microscopy technique for imaging nanosecond decay times of fluorophores. In the case of frequency-domain FLIM, several methods have been described to resolve the relative abundance of two fluorescent species with different fluorescence decay times. Thus far, single-frequency FLIM methods generally have been limited to quantifying two species with monoexponential decay. However, multiexponential decays are the norm rather than the exception, especially for fluorescent proteins and biological samples. Here, we describe a novel method for determining the fractional contribution in each pixel of an image of a sample containing two (multiexponentially) decaying species using single-frequency FLIM. We demonstrate that this technique allows the unmixing of binary mixtures of two spectrally identical cyan or green fluorescent proteins, each with multiexponential decay. Furthermore, because of their spectral identity, quantitative images of the relative molecular abundance of these fluorescent proteins can be generated that are independent of the microscope light path. The method is rigorously tested using samples of known composition and applied to live cell microscopy using cells expressing multiple (multiexponentially decaying) fluorescent proteins.
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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.