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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

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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