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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/19668

Title: Fluorescence recovery after photobleaching in material and life sciences: putting theory into practice
Authors: Loren, Niklas
Hagman, Joel
Jonasson, Jenny K.
Deschout, Hendrik
Bernin, Diana
Cella-Zanacchi, Francesca
Diaspro, Alberto
McNally, James G.
Ameloot, Marcel
Smisdom, Nick
Nyden, Magnus
Hermansson, Anne-Marie
Rudemo, Mats
Braeckmans, Kevin
Issue Date: 2015
Citation: QUARTERLY REVIEWS OF BIOPHYSICS, 48 (3), p. 323-387
Abstract: 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.
Notes: [Loren, Niklas; Hagman, Joel; Hermansson, Anne-Marie] SP Food & Biosci, SE-40229 Gothenburg, Sweden. [Jonasson, Jenny K.; Rudemo, Mats] Chalmers, Dept Math Sci, SE-41296 Gothenburg, Sweden. [Deschout, Hendrik; Braeckmans, Kevin] Univ Ghent, Lab Gen Biochem & Phys Pharm, Biophoton Imaging Grp, B-9000 Ghent, Belgium. [Deschout, Hendrik; Braeckmans, Kevin] Univ Ghent, Ctr Nano & Biophoton, B-9000 Ghent, Belgium. [Bernin, Diana; Hermansson, Anne-Marie] Chalmers, Dept Chem & Biol Engn, SE-41296 Gothenburg, Sweden. [Cella-Zanacchi, Francesca; Diaspro, Alberto] Ist Italiano Tecnol, Nanophys Dept, I-16163 Genoa, Italy. [McNally, James G.] Helmholtz Ctr Berlin, Inst Soft Matter & Funct Mat, D-12489 Berlin, Germany. [Ameloot, Marcel; Smisdom, Nick] Hasselt Univ, B-3500 Hasselt, Belgium. [Smisdom, Nick] Flemish Inst Technol Res, Environm Risk & Hlth Unit, B-2400 Mol, Belgium. [Nyden, Magnus] Univ S Australia, Ian Wark Res Inst, Adelaide, SA 5001, Australia.
URI: http://hdl.handle.net/1942/19668
DOI: 10.1017/S0033583515000013
ISI #: 000360580400002
ISSN: 0033-5835
Category: A1
Type: Journal Contribution
Validation: ecoom, 2016
Appears in Collections: Research publications

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