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  4. Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) with low background and high count-rate per molecule
 
research article

Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) with low background and high count-rate per molecule

Hassler, K.  
•
Leutenegger, M.  
•
Rigler, P.
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2005
Optics Express

We designed a fluorescence correlation spectroscopy (FCS) system for measurements on surfaces. The system consists of an objective-type total internal reflection fluorescence (TIRF) microscopy setup, adapted to measure FCS. Here, the fluorescence exciting evanescent wave is generated by epi-illumination through the periphery of a high NA oil-immersion objective. The main advantages with respect to conventional FCS systems are an improvement in terms of counts per molecule (cpm) and a high signal to background ratio. This is demonstrated by investigating diffusion as well as binding and release of single molecules on a glass surface. Furthermore, the size and shape of the molecule detection efficiency (MDE) function was calculated, using a wave-vectorial approach and taking into account the influence of the dielectric interface on the emission properties of fluorophores. (c) 2005 Optical Society of America.

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Type
research article
DOI
10.1364/OPEX.13.007415
Web of Science ID

WOS:000232058200018

Author(s)
Hassler, K.  
Leutenegger, M.  
Rigler, P.
Rao, R.  
Rigler, R.  
Gosch, M.
Lasser, T.  
Date Issued

2005

Published in
Optics Express
Volume

13

Issue

19

Start page

7415

End page

7423

Subjects

(170.6280) Medical optics and biotechnology

•

Spectroscopy

•

fluorescence and luminescence

•

(240.6490) Optics at surfaces

•

Spectroscopy

•

surface

•

(300.2530) Spectroscopy

•

Fluorescence

•

laser-induced

•

Spectroscopy

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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Available on Infoscience
October 3, 2006
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/234930
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