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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 18, Iss. 22 — Oct. 25, 2010
  • pp: 22886–22905

Straightforward FRAP for quantitative diffusion measurements with a laser scanning microscope

Hendrik Deschout, Joel Hagman, Sophia Fransson, Jenny Jonasson, Mats Rudemo, Niklas Lorén, and Kevin Braeckmans  »View Author Affiliations


Optics Express, Vol. 18, Issue 22, pp. 22886-22905 (2010)
http://dx.doi.org/10.1364/OE.18.022886


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Abstract

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.

© 2010 OSA

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.1790) Medical optics and biotechnology : Confocal microscopy
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Microscopy

History
Original Manuscript: June 21, 2010
Revised Manuscript: August 20, 2010
Manuscript Accepted: October 7, 2010
Published: October 14, 2010

Virtual Issues
Vol. 6, Iss. 1 Virtual Journal for Biomedical Optics

Citation
Hendrik Deschout, Joel Hagman, Sophia Fransson, Jenny Jonasson, Mats Rudemo, Niklas Lorén, and Kevin Braeckmans, "Straightforward FRAP for quantitative diffusion measurements with a laser scanning microscope," Opt. Express 18, 22886-22905 (2010)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-18-22-22886


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