Fluorescence recovery after photobleaching in material and life sciences: Putting theory into practice
Artikel i vetenskaplig tidskrift, 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.

Författare

Jenny Jonasson

Chalmers, Matematiska vetenskaper, matematisk statistik

Göteborgs universitet

Hendrik Deschout

Universiteit Gent

Diana Bernin

SuMo Biomaterials

Göteborgs universitet

Chalmers, Kemi och kemiteknik, Tillämpad kemi, Teknisk ytkemi

F. Cella-Zanacchi

Istituto Italiano di Tecnologia

A. Diaspro

Istituto Italiano di Tecnologia

J.G. McNally

M. Ameloot

Universiteit Hasselt

N. Smisdom

Flemish Institute for Technological Research

Universiteit Hasselt

Magnus Nydén

University of South Australia

Anne-Marie Hermansson

SuMo Biomaterials

Chalmers, Biologi och bioteknik, Livsmedelsvetenskap

Mats Rudemo

Göteborgs universitet

SuMo Biomaterials

Chalmers, Matematiska vetenskaper, matematisk statistik

Kevin Braeckmans

Universiteit Gent

Quarterly Reviews of Biophysics

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

Vol. 48 323-387

Ämneskategorier

Fysik

Materialkemi

Annan matematik

Annan fysik

Fundament

Grundläggande vetenskaper

Styrkeområden

Livsvetenskaper och teknik

Materialvetenskap

DOI

10.1017/S0033583515000013