Fluorescence recovery after photobleaching in material and life sciences: Putting theory into practice
Journal article, 2015

Copyright © 2015 Cambridge University Press.Fluorescence recovery after photobleaching (FRAP) is a versatile tool for determining diffusion and interaction/binding properties in biological and material sciences. An understanding of the mechanisms controlling the diffusion requires a deep understanding of structure-interaction-diffusion relationships. In cell biology, for instance, this applies to the movement of proteins and lipids in the plasma membrane, cytoplasm and nucleus. In industrial applications related to pharmaceutics, foods, textiles, hygiene products and cosmetics, the diffusion of solutes and solvent molecules contributes strongly to the properties and functionality of the final product. All these systems are heterogeneous, and accurate quantification of the mass transport processes at the local level is therefore essential to the understanding of the properties of soft (bio)materials. FRAP is a commonly used fluorescence microscopy-based technique to determine local molecular transport at the micrometer scale. A brief high-intensity laser pulse is locally applied to the sample, causing substantial photobleaching of the fluorescent molecules within the illuminated area. This causes a local concentration gradient of fluorescent molecules, leading to diffusional influx of intact fluorophores from the local surroundings into the bleached area. Quantitative information on the molecular transport can be extracted from the time evolution of the fluorescence recovery in the bleached area using a suitable model. A multitude of FRAP models has been developed over the years, each based on specific assumptions. This makes it challenging for the non-specialist to decide which model is best suited for a particular application. Furthermore, there are many subtleties in performing accurate FRAP experiments. For these reasons, this review aims to provide an extensive tutorial covering the essential theoretical and practical aspects so as to enable accurate quantitative FRAP experiments for molecular transport measurements in soft (bio)materials.

Author

Jenny Jonasson

Chalmers, Mathematical Sciences, Mathematical Statistics

University of Gothenburg

Hendrik Deschout

Ghent university

Diana Bernin

Chalmers, Chemistry and Chemical Engineering, Applied Chemistry, Applied Surface Chemistry

University of Gothenburg

SuMo Biomaterials

F. Cella-Zanacchi

Istituto Italiano di Tecnologia

A. Diaspro

Istituto Italiano di Tecnologia

J.G. McNally

M. Ameloot

Universiteit Hasselt

N. Smisdom

Universiteit Hasselt

Flemish Institute for Technological Research

Magnus Nydén

University of South Australia

Anne-Marie Hermansson

SuMo Biomaterials

Chalmers, Biology and Biological Engineering, Food and Nutrition Science

Mats Rudemo

Chalmers, Mathematical Sciences, Mathematical Statistics

SuMo Biomaterials

University of Gothenburg

Kevin Braeckmans

Ghent university

Quarterly Reviews of Biophysics

0033-5835 (ISSN) 1469-8994 (eISSN)

Vol. 48 3 323-387

Subject Categories

Physical Sciences

Materials Chemistry

Other Mathematics

Other Physics Topics

Roots

Basic sciences

Areas of Advance

Life Science Engineering (2010-2018)

Materials Science

DOI

10.1017/S0033583515000013

More information

Latest update

11/30/2018