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Rectangle FRAP for measuring diffusion with a laser scanning microscope

Ranhua Xiong (UGent) , Hendrik Deschout (UGent) , Jo Demeester (UGent) , Stefaan De Smedt (UGent) and Kevin Braeckmans (UGent)
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Center for nano- and biophotonics (NB-Photonics)
Abstract
Fluorescence recovery after photobleaching (FRAP) is one of the most useful microscopy techniques for studying the mobility of molecules in terms of a diffusion coefficient. Here, we describe a FRAP method that allows such measurements, relying on the photobleaching of a rectangular region of any size and aspect ratio. We start with a brief overview of the rectangle FRAP theory and next we provide guidelines for performing FRAP measurements, including a discussion of the experimental set-up and the data analysis. Finally, we discuss how to verify correct use of the rectangle FRAP method using test solutions.
Keywords
Fluorescence microscopy, Confocal laser scanning microscopy, Fluorescence recovery after photobleaching, Diffusion coefficient

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Citation

Please use this url to cite or link to this publication:

Chicago
Xiong, Ranhua, Hendrik Deschout, Jo Demeester, Stefaan De Smedt, and Kevin Braeckmans. 2013. “Rectangle FRAP for Measuring Diffusion with a Laser Scanning Microscope.” In Fluorescence Spectroscopy and Microscopy : Methods and Protocols, ed. Yves Engelborghs and Antonie JWG Visser, 1076:433–441. New York, NY, USA: Humana Press.
APA
Xiong, R., Deschout, H., Demeester, J., De Smedt, S., & Braeckmans, K. (2013). Rectangle FRAP for measuring diffusion with a laser scanning microscope. In Y. Engelborghs & A. J. Visser (Eds.), Fluorescence spectroscopy and microscopy : methods and protocols (Vol. 1076, pp. 433–441). New York, NY, USA: Humana Press.
Vancouver
1.
Xiong R, Deschout H, Demeester J, De Smedt S, Braeckmans K. Rectangle FRAP for measuring diffusion with a laser scanning microscope. In: Engelborghs Y, Visser AJ, editors. Fluorescence spectroscopy and microscopy : methods and protocols. New York, NY, USA: Humana Press; 2013. p. 433–41.
MLA
Xiong, Ranhua, Hendrik Deschout, Jo Demeester, et al. “Rectangle FRAP for Measuring Diffusion with a Laser Scanning Microscope.” Fluorescence Spectroscopy and Microscopy : Methods and Protocols. Ed. Yves Engelborghs & Antonie JWG Visser. Vol. 1076. New York, NY, USA: Humana Press, 2013. 433–441. Print.
@incollection{4097329,
  abstract     = {Fluorescence recovery after photobleaching (FRAP) is one of the most useful microscopy techniques for studying the mobility of molecules in terms of a diffusion coefficient. Here, we describe a FRAP method that allows such measurements, relying on the photobleaching of a rectangular region of any size and aspect ratio. We start with a brief overview of the rectangle FRAP theory and next we provide guidelines for performing FRAP measurements, including a discussion of the experimental set-up and the data analysis. Finally, we discuss how to verify correct use of the rectangle FRAP method using test solutions.},
  author       = {Xiong, Ranhua and Deschout, Hendrik and Demeester, Jo and De Smedt, Stefaan and Braeckmans, Kevin},
  booktitle    = {Fluorescence spectroscopy and microscopy : methods and protocols},
  editor       = {Engelborghs, Yves and Visser, Antonie JWG},
  isbn         = {9781627036481},
  issn         = {1064-3745},
  language     = {eng},
  pages        = {433--441},
  publisher    = {Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Rectangle FRAP for measuring diffusion with a laser scanning microscope},
  url          = {http://dx.doi.org/10.1007/978-1-62703-649-8\_18},
  volume       = {1076},
  year         = {2013},
}

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