Straightforward FRAP for quantitative diffusion measurements with a laser scanning microscope
Journal article, 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

Author

Hendrik Deschout

Ghent university

Joel H Hagman

Chalmers, Chemical and Biological Engineering, Applied Surface Chemistry

SuMo Biomaterials

Sophia Fransson

Chalmers, Chemical and Biological Engineering, Applied Surface Chemistry

Jenny Jonasson

SuMo Biomaterials

University of Gothenburg

Chalmers, Mathematical Sciences, Mathematical Statistics

Mats Rudemo

University of Gothenburg

Chalmers, Mathematical Sciences, Mathematical Statistics

Niklas Lorén

Chalmers, Chemical and Biological Engineering, Applied Surface Chemistry

SuMo Biomaterials

Kevin Braeckmans

Ghent university

Optics Express

1094-4087 (ISSN)

Vol. 18 22 22886-22905

Subject Categories

Medical Laboratory and Measurements Technologies

Atom and Molecular Physics and Optics

Areas of Advance

Life Science Engineering (2010-2018)

Materials Science

DOI

10.1364/OE.18.022886

More information

Latest update

3/6/2018 1