FRAP Analysis: Accounting for Bleaching during Image Capture
Open Access
- 9 August 2012
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 7 (8) , e42854
Abstract
The analysis of Fluorescence Recovery After Photobleaching (FRAP) experiments involves mathematical modeling of the fluorescence recovery process. An important feature of FRAP experiments that tends to be ignored in the modeling is that there can be a significant loss of fluorescence due to bleaching during image capture. In this paper, we explicitly include the effects of bleaching during image capture in the model for the recovery process, instead of correcting for the effects of bleaching using reference measurements. Using experimental examples, we demonstrate the usefulness of such an approach in FRAP analysis.Keywords
This publication has 25 references indexed in Scilit:
- Monitoring Dynamic Binding of Chromatin Proteins In Vivo by Fluorescence Recovery After Photobleaching2011
- Fluorescence recovery after photobleaching on the confocal laser-scanning microscope: generalized model without restriction on the size of the photobleached diskJournal of Biomedical Optics, 2011
- Straightforward FRAP for quantitative diffusion measurements with a laser scanning microscopeOptics Express, 2010
- FRAP and kinetic modeling in the analysis of nuclear protein dynamics: what do we really know?Current Opinion in Cell Biology, 2010
- In vitro FRAP reveals the ATP-dependent nuclear mobilization of the exon junction complex protein SRm160Journal of Cell Biology, 2004
- Measurement of Dynamic Protein Binding to Chromatin In Vivo, Using Photobleaching Microscopy2003
- Kinetic modelling approaches to in vivo imagingNature Reviews Molecular Cell Biology, 2001
- FRAP reveals that mobility of oestrogen receptor-α is ligand- and proteasome-dependentNature Cell Biology, 2000
- Measurement of mass transport and reaction parameters in bulk solution using photobleaching. Reaction limited binding regimeBiophysical Journal, 1991
- Quantification of transport and binding parameters using fluorescence recovery after photobleaching. Potential for in vivo applicationsBiophysical Journal, 1990