ArticleBiophysical journal2008
A new FRAP/FRAPa method for three-dimensional diffusion measurements based on multiphoton excitation 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 27 papers.
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Who cites it
27 citing papers in PubMed.
- Exploring focal adhesion data: dynamic parameter extraction from FRAP and FLAP experiments using chemical master equation.Frontiers in molecular biosciences · 2025Article
- Selective activation of photoactivatable fluorescent protein based on binary holography.Biomedical optics express · 2024Article
- What's past is prologue: FRAP keeps delivering 50 years later.Biophysical journal · 2023Review
- Beyond analytic solution: Analysis of FRAP experiments by spatial simulation of the forward problem.Biophysical journal · 2023Article
- Expanding the applicability of multiphoton fluorescence recovery after photobleaching by incorporating shear stress in laminar flow.Journal of biomedical optics · 2023Article
- A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells.STAR protocols · 2021Article
- Parameter estimation in fluorescence recovery after photobleaching: quantitative analysis of protein binding reactions and diffusion.Journal of mathematical biology · 2021Article
- A noninvasive fluorescence imaging-based platform measures 3D anisotropic extracellular diffusion.Nature communications · 2021Article
- Methods for characterizing the material properties of biomolecular condensates.Methods in enzymology · 2021Article
- Back to the Future: Genetically Encoded Fluorescent Proteins as Inert Tracers of the Intracellular Environment.International journal of molecular sciences · 2020Review
- Quantifying Dynamics in Phase-Separated Condensates Using Fluorescence Recovery after Photobleaching.Biophysical journal · 2019Article
- Article
- Measurement of Rapid Protein Diffusion in the Cytoplasm by Photo-Converted Intensity Profile Expansion.Cell reports · 2017Article
- Improving Parameter Inference from FRAP Data: an Analysis Motivated by Pattern Formation in the Drosophila Wing Disc.Bulletin of mathematical biology · 2017Article
- Single- and two-photon fluorescence recovery after photobleaching.Cold Spring Harbor protocols · 2015Article
- Phototransformable fluorescent proteins: which one for which application?Histochemistry and cell biology · 2014Review
- Measurement of three-dimensional anisotropic diffusion by multiphoton fluorescence recovery after photobleaching.Annals of biomedical engineering · 2014Article
- Molecular diffusion and binding analyzed with FRAP.Protoplasma · 2014Review
- Presynaptic alpha-synuclein aggregation in a mouse model of Parkinson's disease.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2014Article
- FRAP in pharmaceutical research: practical guidelines and applications in drug delivery.Pharmaceutical research · 2014Review
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Authors and funding
8 authors.
Funding
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Abstract
We present a new convenient method for quantitative three-dimensionally resolved diffusion measurements based on the photobleaching (FRAP) or photoactivation (FRAPa) of a disk-shaped area by the scanning laser beam of a multiphoton microscope. Contrary to previously reported spot-photobleaching protocols, this method has the advantage of full scalability of the size of the photobleached area and thus the range of diffusion coefficients, which can be measured conveniently. The method is compatible with low as well as high numerical aperture objective lenses, allowing us to perform quantitative diffusion measurements in three-dimensional extended samples as well as in very small volumes, such as cell nuclei. Furthermore, by photobleaching/photoactivating a large area, diffusion along the optical axis can be measured separately, which is convenient when studying anisotropic diffusion. First, we show the rigorous mathematical derivation of the model, leading to a closed-form formula describing the fluorescence recovery/redistribution phase. Next, the ability of the multiphoton FRAP method to correctly measure absolute diffusion coefficients is tested thoroughly on many test solutions of FITC-dextrans covering a wide range of diffusion coefficients. The same is done for the FRAPa method on a series of photoactivatable green fluorescent protein solutions with different viscosities. Finally, we apply the method to photoactivatable green fluorescent protein diffusing freely in the nucleus of living NIH-3T3 mouse embryo fibroblasts.
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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.