ArticleBiophysical journal2012
Live-cell fluorescence microscopy with molecular biosensors: what are we really measuring?
Article in Biophysical journal, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 35 citations in OpenAlex.
- On the inference of ERK signaling dynamics from protein biosensor measurements.Molecular biology of the cell · 2023Article
- Device Processing Challenges for Miniaturized Sensing Systems Targeting Biological Fluids.Biomedical materials & devices (New York, N.Y.) · 2022Review
- Article
- Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Design and evaluation of engineered protein biosensors for live-cell imaging of EGFR phosphorylation.Science signaling · 2019Article
- Single-Cell Imaging of Metastatic Potential of Cancer Cells.iScience · 2018Article
- Article
- Radiative decay engineering 8: Coupled emission microscopy for lens-free high-throughput fluorescence detection.Analytical biochemistry · 2017Article
- A Fluoro-Chromogenic Sensor Based on Organic Molecular Framework for CuJournal of fluorescence · 2017Review
- Are Filopodia Privileged Signaling Structures in Migrating Cells?Biophysical journal · 2016Article
- Next-generation RNA-based fluorescent biosensors enable anaerobic detection of cyclic di-GMP.Nucleic acids research · 2016Article
- Using fluctuation analysis to establish causal relations between cellular events without experimental perturbation.Biophysical journal · 2014Review
- Deactivation of a negative regulator: a distinct signal transduction mechanism, pronounced in Akt signaling.Biophysical journal · 2014Article
- Equilibrium and dynamic design principles for binding molecules engineered for reagentless biosensors.Analytical biochemistry · 2014Article
- Biosensor architectures for high-fidelity reporting of cellular signaling.Biophysical journal · 2014Article
- Glutamine flux imaging using genetically encoded sensors.Journal of visualized experiments : JoVE · 2014Article
- Microfluidic probe for single-cell analysis in adherent tissue culture.Nature communications · 2014Article
- Development of FRET biosensors for mammalian and plant systems.Protoplasma · 2014Review
- Monitoring of post-translational modification dynamics with genetically encoded fluorescent reporters.Biopolymers · 2014Review
- Data-driven modeling reconciles kinetics of ERK phosphorylation, localization, and activity states.Molecular systems biology · 2014Article
Corrections and comments
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Authors and funding
1 author at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Engineered protein biosensors, such as those based on Förster resonance energy transfer, membrane translocation, or solvatochromic shift, are being used in combination with live-cell fluorescence microscopy to reveal kinetics and spatial localization of intracellular processes as they occur. Progress in the application of this approach has been steady, yet its general suitability for quantitative measurements remains unclear. To address the pitfalls of the biosensor approach in quantitative terms, simple reaction-diffusion models were analyzed. The analysis shows that although high-affinity molecular recognition allows robust detection of the fluorescence readout, either of two detrimental effects is fostered. Binding of an intramolecular biosensor or of a relatively abundant intermolecular biosensor introduces observer effects in which the dynamics of the target molecule under study are significantly perturbed, whereas binding of a sparingly expressed intermolecular biosensor is subject to a saturation effect, where the pool of unbound biosensor is significantly depleted. The analysis explores how these effects are manifest in the kinetics and spatial gradients of the biosensor-target complex. A sobering insight emerges: the observer or saturation effect is always significant; the question is whether or not it can be tolerated or accounted for. The challenge in managing the adverse effects is that specification of the biosensor-target affinity to within a certain order of magnitude is required.
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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.