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research article

High light field confinement for Fluorescence Correlation Spectroscopy using a Solid Immersion Lens

Serov, A.  
•
Rao, R.  
•
Goesch, M.
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2004
Biosensors and Bioelectronics

In this paper we present recent single molecule detection experiment using a solid immersion lens (SIL) for fluorescent correlation spectroscopy measurements. We compared the performance of the SIL in combination with an air objective (40×, numerical aperture (NA)=0.6) with a water immersion objective (40×, NA=1.15) in a confocal microscope system (ConfoCorr 1). Important parameters for single molecule experiments such as collection efficiency and excitation field confinement were investigated. Although the two set-ups have similar numerical aperture the measurements demonstrated higher field confinement and better collection efficiency for the SIL system in comparison to the conventional confocal set-up. Adding spherical aberrations shifts the sample volume up to 4 μm away from the plane surface of the SIL and conserves a diffraction limited focal volume. In this case the FCS autocorrelation demonstrates a free 3D diffusion of dye molecules in a highly confined light field.

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Type
research article
DOI
10.1016/j.bios.2004.02.028
Author(s)
Serov, A.  
Rao, R.  
Goesch, M.
Anhut, T.  
Martin, D.
Brunner, R.
Rigler, P.
Lasser, T.  
Date Issued

2004

Publisher

Elsevier

Published in
Biosensors and Bioelectronics
Volume

20

Issue

3

Start page

431

End page

435

Subjects

Fluorescent correlation spectroscopy

•

Solid immersion lens

•

Single molecule detection

•

High resolution

Note

Institute for Biomedical Imaging, Optics, and Engineering of the Swiss Federal Institute of Technology (EPFL), CH-1015, Lausanne, Switzerland Department of Medical Biochemistry and Biophysics, Karolinska Institute, S-171 77, Stockholm, Sweden Carl Zeiss Jena GmbH, Germany

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October 3, 2006
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/234923
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