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• W1526551292 abstract "Confocal fluorescence recovery after photobleaching (FRAP) is today the prevalent tool when studying the diffusional and kinetic properties of proteins in living cells. Obtaining quantitative data for diffusion coefficients via FRAP, however, is challenged by the fact that both bleaching and scanning take a finite time. Starting from an experimental case, it is shown by means of computer simulations that this intrinsic temporal limitation can lead to a gross underestimation of diffusion coefficients. Determining the binding kinetics of proteins to membranes with FRAP is further shown to be severely hampered by additional diffusional contributions, e.g. diffusion-limited binding. In some cases, the binding kinetics may even be masked entirely by diffusion. As current efforts to approach biological problems with biophysical models have to rely on experimentally determined model parameters, e.g. binding rates and diffusion constants, it is proposed that the accuracy in evaluating FRAP measurements can be improved by means of accompanying computer simulations." @default.
• W1526551292 created "2016-06-24" @default.
• W1526551292 creator A5054742160 @default.
• W1526551292 date "2004-07-26" @default.
• W1526551292 modified "2023-09-25" @default.
• W1526551292 title "Challenges and Artifacts in Quantitative Photobleaching Experiments" @default.
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• W1526551292 doi "https://doi.org/10.1111/j.1600-0854.2004.00215.x" @default.
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