ReviewAmerican journal of physiology. Cell physiology2015
A practical method for monitoring FRET-based biosensors in living animals using two-photon microscopy.
Review in American journal of physiology. Cell physiology, 2015. 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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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
21 citing papers in PubMed, 29 citations in OpenAlex.
- Expanding Horizons in Advancements of FRET Biosensing Technologies.Biosensors · 2025Review
- Fluorescent sensors for intracellular signaling in the brain: imaging neurons, glia, and vascular cells.Neurophotonics · 2025Article
- Cell-cell communication: new insights and clinical implications.Signal transduction and targeted therapy · 2024Review
- Opioid-Modulated Receptor Localization and Erk1/2 Phosphorylation in Cells Coexpressing μ-Opioid and Nociceptin Receptors.International journal of molecular sciences · 2023Article
- Quantifying molecular- to cellular-level forces in living cells.Journal of physics D: Applied physics · 2021Article
- Genetically encoded sensors towards imaging cAMP and PKA activity in vivo.Journal of neuroscience methods · 2021Review
- The chemical tools for imaging dopamine release.Cell chemical biology · 2021Review
- The Indiana O'Brien Center for Advanced Renal Microscopic Analysis.American journal of physiology. Renal physiology · 2021Review
- Evaluation of Caspase-3 Activity During Apoptosis with Fluorescence Lifetime-Based Cytometry Measurements and Phasor Analyses.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2020Article
- Milestones in the development and implementation of FRET-based sensors of intracellular signals: A biological perspective of the history of FRET.Cellular signalling · 2020Review
- Dopamine D1 receptor signalling in dyskinetic Parkinsonian rats revealed by fiber photometry using FRET-based biosensors.Scientific reports · 2020Article
- Illuminating the kinome: Visualizing real-time kinase activity in biological systems using genetically encoded fluorescent protein-based biosensors.Current opinion in chemical biology · 2020Review
- A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo.Scientific reports · 2019Article
- Advanced FRET normalization allows quantitative analysis of protein interactions including stoichiometries and relative affinities in living cells.Scientific reports · 2019Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- Imaging the Cardiac Extracellular Matrix.Advances in experimental medicine and biology · 2018Review
- Imaging sympathetic neurogenic CaAutonomic neuroscience : basic & clinical · 2017Review
- Intravital microscopy of biosensor activities and intrinsic metabolic states.Methods (San Diego, Calif.) · 2017Article
- Surgical preparation of rats and mice for intravital microscopic imaging of abdominal organs.Methods (San Diego, Calif.) · 2017Article
- A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy.Nature protocols · 2016Article
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9 authors at 5 institutions in 1 country.
Funding
Abstract
The commercial availability of multiphoton microscope systems has nurtured the growth of intravital microscopy as a powerful technique for evaluating cell biology in the relevant context of living animals. In parallel, new fluorescent protein (FP) biosensors have become available that enable studies of the function of a wide range of proteins in living cells. Biosensor probes that exploit Förster resonance energy transfer (FRET) are among the most sensitive indicators of an array of cellular processes. However, differences between one-photon and two-photon excitation (2PE) microscopy are such that measuring FRET by 2PE in the intravital setting remains challenging. Here, we describe an approach that simplifies the use of FRET-based biosensors in intravital 2PE microscopy. Based on a systematic comparison of many different FPs, we identified the monomeric (m) FPs mTurquoise and mVenus as particularly well suited for intravital 2PE FRET studies, enabling the ratiometric measurements from linked FRET probes using a pair of experimental images collected simultaneously. The behavior of the FPs is validated by fluorescence lifetime and sensitized emission measurements of a set of FRET standards. The approach is demonstrated using a modified version of the AKAR protein kinase A biosensor, first in cells in culture, and then in hepatocytes in the liver of living mice. The approach is compatible with the most common 2PE microscope configurations and should be applicable to a variety of different FRET probes.
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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.