Pixel-based analysis of FRAP data with a general initial bleaching profile

Journal of Microscopy
J K JonassonM Rudemo

Abstract

In Jonasson et al. (2008), we presented a new pixel-based maximum likelihood framework for the estimation of diffusion coefficients from data on fluorescence recovery after photobleaching (FRAP) with confocal laser scanning microscopy (CLSM). The main method there, called the Gaussian profile method below, is based on the assumption that the initial intensity profile after photobleaching is approximately Gaussian. In the present paper, we introduce a method, called the Monotone profile method, where the maximum likelihood framework is extended to a general initial bleaching profile only assuming that the profile is a non-decreasing function of the distance to the bleaching centre. The statistical distribution of the image noise is further assumed to be Poisson instead of normal, which should be a more realistic description of the noise in the detector. The new Monotone profile method and the Gaussian profile method are applied to FRAP data on swelling of super absorbent polymers (SAP) in water with a Fluorescein probe. The initial bleaching profile is close to a step function at low degrees of swelling and close to a Gaussian profile at high degrees of swelling. The results obtained from the analysis of the FRAP data are corrob...Continue Reading

References

Jan 1, 1972·Journal of Clinical Pathology. Supplement (Royal College of Pathologists)·R M Barlow
Jun 2, 2009·Advances in Colloid and Interface Science·Niklas LorénAnne-Marie Hermansson

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Citations

Sep 11, 2013·Pharmaceutical Research·Hendrik DeschoutKevin Braeckmans
Nov 5, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Nadia BourouinaJ Mieke Kleijn
Sep 1, 2015·Quarterly Reviews of Biophysics·Niklas LorénKevin Braeckmans
Nov 29, 2020·Journal of Microscopy·Victor Wåhlstrand SkärströmMagnus Röding
Dec 20, 2017·The Journal of Physical Chemistry. B·David SustrDmitry Volodkin

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