Derivation of a closed form analytical expression for fluorescence recovery after photo bleaching in the case of continuous bleaching during read out

Measurements of very slow diffusive processes in membranes, like the diffusion of integral membrane proteins, by fluorescence recovery after photo bleaching (FRAP) are hampered by bleaching of the probe during the read out of the fluorescence recovery. In the limit of long observation time (very slo...

Full description

Saved in:
Bibliographic Details
Published in:The European physical journal. E, Soft matter and biological physics Soft matter and biological physics, 2005, Vol.16 (1), p.81-87
Main Authors: ENDRESS, E, WEIGELT, S, REENTS, G, BAYERL, T. M
Format: Article
Language:English
Subjects:
Animals
Cell Membrane - chemistry
Cells, Cultured
Chemistry
Colloidal state and disperse state
Computer Simulation
Cytochrome P-450 Enzyme System - analysis
Cytochrome P-450 Enzyme System - chemistry
Diffusion
Exact sciences and technology
Fluorescence Recovery After Photobleaching - methods
General and physical chemistry
Membrane Fluidity
Membrane Proteins - analysis
Membrane Proteins - chemistry
Membranes
Microsomes, Liver - chemistry
Models, Biological
Models, Chemical
Rabbits
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Measurements of very slow diffusive processes in membranes, like the diffusion of integral membrane proteins, by fluorescence recovery after photo bleaching (FRAP) are hampered by bleaching of the probe during the read out of the fluorescence recovery. In the limit of long observation time (very slow diffusion as in the case of large membrane proteins), this bleaching may cause errors to the recovery function and thus provides error-prone diffusion coefficients. In this work we present a new approach to a two-dimensional closed form analytical solution of the reaction-diffusion equation, based on the addition of a dissipative term to the conventional diffusion equation. The calculation was done assuming (i) a Gaussian laser beam profile for bleaching the spot and (ii) that the fluorescence intensity profile emerging from the spot can be approximated by a two-dimensional Gaussian. The detection scheme derived from the analytical solution allows for diffusion measurements without the constraint of observation bleaching. Recovery curves of experimental FRAP data obtained under non-negligible read-out bleaching for native membranes (rabbit endoplasmic reticulum) on a planar solid support showed excellent agreement with the analytical solution and allowed the calculation of the lipid diffusion coefficient.
ISSN:1292-8941
1292-895X
DOI:10.1140/epje/e2005-00010-5