Straightforward FRAP for quantitative diffusion measurements with a laser scanning microscope
Artikel i vetenskaplig tidskrift, 2010
Confocal or multi-photon laser scanning microscopes are convenient tools to perform FRAP diffusion measurements. Despite its popularity, accurate FRAP remains often challenging since current methods are either limited to relatively large bleach regions or can be complicated for non-specialists. In order to bring reliable quantitative FRAP measurements to the broad community of laser scanning microscopy users, here we have revised FRAP theory and present a new pixel based FRAP method relying on the photo bleaching of rectangular regions of any size and aspect ratio. The method allows for fast and straightforward quantitative diffusion measurements due to a closed–form expression for the recovery process utilizing all available spatial and temporal data. After a detailed validation, its versatility is demonstrated by diffusion studies in heterogeneous biopolymer mixtures.
Fluorescence microscopy
Medical optics and biotechnology
Confocal microscopy
Microscopy
Författare
Hendrik Deschout
Universiteit Gent
Joel H Hagman
SuMo Biomaterials
Chalmers, Kemi- och bioteknik, Teknisk ytkemi
Sophia Fransson
Chalmers, Kemi- och bioteknik, Teknisk ytkemi
Jenny Jonasson
SuMo Biomaterials
Chalmers, Matematiska vetenskaper, Matematisk statistik
Göteborgs universitet
Mats Rudemo
Göteborgs universitet
Chalmers, Matematiska vetenskaper, Matematisk statistik
Niklas Lorén
Chalmers, Kemi- och bioteknik, Teknisk ytkemi
SuMo Biomaterials
Kevin Braeckmans
Universiteit Gent
Optics Express
1094-4087 (ISSN) 10944087 (eISSN)
Vol. 18 22 22886-22905Ämneskategorier
Medicinsk laboratorie- och mätteknik
Atom- och molekylfysik och optik
Styrkeområden
Livsvetenskaper och teknik (2010-2018)
Materialvetenskap
DOI
10.1364/OE.18.022886