Abstract
Fluorescence recovery after photobleaching (FRAP) is used for the study of the diffusion and binding of target proteins in biological cells. But the approach poses problems in analytical tractability of measurements in an inhomogeneous medium. Here, we present a method to solve the problem applying an integral transform of the kinetics of the entire cellular two-dimensional spatial pattern of recovered fluorescence. The experimental data were obtained with the Leica TCS SP5X confocal microscope. The method was demonstrated by the evaluation of the association and the dissociation rate constants and the diffusion coefficient of GFP-tagged heterochromatin protein 1 and GFP-tagged c-Myc protein in the nuclei of mouse embryonic fibroblasts. The main novelty of the approach is that it takes into account the inhomogeneous distribution of binding sites and does not require the complete mathematical solution of a corresponding system of diffusion-reaction equations that is typical for conventional FRAP data processing.This work was supported by the Russian Ministry of Education and Science (P1039, 14.740.11.1194, 14.740.11.0921), by the European Union project (COST TD09/05) and Marie Curie project PIRSES-GA-2010-269156-LCS, by the Grant Agency of the Czech Republic (P302/10/1022). The work also has been supported by the Academy of Sciences of the Czech Republic, and by the Ministry of Education Youth and Sports of the Czech Republic: the research projects Nos. LC535, LC06027, ME 919, AVOZ50040702, AVOZ50040507, and COST-CZ LC11020.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
