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

Microscopy

Medical optics and biotechnology

Confocal microscopy

Författare

Hendrik Deschout

Universiteit Gent

Joel H Hagman

Chalmers, Kemi- och bioteknik, Teknisk ytkemi

SuMo Biomaterials

Sophia Fransson

Chalmers, Kemi- och bioteknik, Teknisk ytkemi

Jenny Jonasson

SuMo Biomaterials

Göteborgs universitet

Chalmers, Matematiska vetenskaper, matematisk statistik

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)

Vol. 18 22886-22905

Ämneskategorier

Medicinsk laboratorie- och mätteknik

Atom- och molekylfysik och optik

Styrkeområden

Livsvetenskaper och teknik

Materialvetenskap

DOI

10.1364/OE.18.022886