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
Medical optics and biotechnology
Confocal microscopy
Microscopy
Author
Hendrik Deschout
Ghent university
Joel H Hagman
SuMo Biomaterials
Chalmers, Chemical and Biological Engineering, Applied Surface Chemistry
Sophia Fransson
Chalmers, Chemical and Biological Engineering, Applied Surface Chemistry
Jenny Jonasson
SuMo Biomaterials
Chalmers, Mathematical Sciences, Mathematical Statistics
University of Gothenburg
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) 10944087 (eISSN)
Vol. 18 22 22886-22905Subject 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