ArticlePloS one2015
Validation of Normalizations, Scaling, and Photofading Corrections for FRAP Data Analysis.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
What it found
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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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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
20 citing papers in PubMed, 38 citations in OpenAlex.
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- PIM1 controls GBP1 activity to limit self-damage and to guard against pathogen infection.Science (New York, N.Y.) · 2023Article
- Probing Microscale Structuring-Induced Phase Separation with Fluorescence Recovery Diffusion Dynamics in Poly(ethylene glycol) Solutions.ACS omega · 2023Article
- What's past is prologue: FRAP keeps delivering 50 years later.Biophysical journal · 2023Review
- Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer.Nature methods · 2023Article
- Cell-mimicking polyethylene glycol-diacrylate based nanolipogel for encapsulation and delivery of hydrophilic biomolecule.Frontiers in bioengineering and biotechnology · 2023Article
- Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction-Associated Polarity Proteins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- MyD88 oligomer size functions as a physical threshold to trigger IL1R Myddosome signaling.The Journal of cell biology · 2021Article
- Apoptosis-inducing anti-HER2 agents operate through oligomerization-induced receptor immobilization.Communications biology · 2021Article
- A Simple Derivation of Diffusion Fluorescence Correlation Spectroscopy Equations.Journal of fluorescence · 2020Article
- A Highly Accurate Pixel-Based FRAP Model Based on Spectral-Domain Numerical Methods.Biophysical journal · 2019Article
- Analysis of Active Transport by Fluorescence Recovery after Photobleaching.Biophysical journal · 2017Article
- FRAP to Characterize Molecular Diffusion and Interaction in Various Membrane Environments.PloS one · 2016Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescence Recovery After Photobleaching (FRAP) has been a versatile tool to study transport and reaction kinetics in live cells. Since the fluorescence data generated by fluorescence microscopy are in a relative scale, a wide variety of scalings and normalizations are used in quantitative FRAP analysis. Scaling and normalization are often required to account for inherent properties of diffusing biomolecules of interest or photochemical properties of the fluorescent tag such as mobile fraction or photofading during image acquisition. In some cases, scaling and normalization are also used for computational simplicity. However, to our best knowledge, the validity of those various forms of scaling and normalization has not been studied in a rigorous manner. In this study, we investigate the validity of various scalings and normalizations that have appeared in the literature to calculate mobile fractions and correct for photofading and assess their consistency with FRAP equations. As a test case, we consider linear or affine scaling of normal or anomalous diffusion FRAP equations in combination with scaling for immobile fractions. We also consider exponential scaling of either FRAP equations or FRAP data to correct for photofading. Using a combination of theoretical and experimental approaches, we show that compatible scaling schemes should be applied in the correct sequential order; otherwise, erroneous results may be obtained. We propose a hierarchical workflow to carry out FRAP data analysis and discuss the broader implications of our findings for FRAP data analysis using a variety of kinetic models.
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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.