Special Section on Mediterranean Nanophotonics

Tethered particle motion mediated by scattering from gold nanoparticles and darkfield microscopy

[+] Author Affiliations
Heidelinde R. C. Dietrich, Bernd Rieger

Department of Imaging Science and Technology, Delft University of Technology, Lorentzweg 1, Delft, ZH 2628 CJ Netherlands

Frank G. M. Wiertz, Frederik H. De Groote, Hendrik A. Heering

Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, Leiden, 2333 CC Netherlands

Ian T. Young, Yuval Garini

Department of Imaging Science and Technology, Delft University of Technology, Lorentzweg 1, Delft, ZH 2628 CJ Netherlands

J. Nanophoton. 3(1), 031795 (June 22, 2009). doi:10.1117/1.3174445
History: Received March 23, 2009; Revised June 11, 2009; Accepted June 17, 2009; June 22, 2009; Online June 22, 2009
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Abstract

We measured light scattered from gold nanoparticles with darkfield microscopy in order to perform single molecule detection based on tethered particle motion (TPM). This combination results in a signal to noise ratio of about 40 dB, which allowed us to use 80 nm diameter gold particles as reporters instead of the typically used polystyrene particles whose sizes are up to 1 μm. The particle size is crucial in TPM experiments as it can induce a volume-exclusion effect, which results in a stretching force acting on the DNA tether. This affects both the biophysical and statistical properties of the anchored DNA and hence the interpretation of the experimental data. We demonstrated that the gold nanoparticles and darkfield microscopy can be used to characterize the confined Brownian motion of dsDNA-tethered gold particles with a spatial precision of 3 nm. Physical parameters such as the spring constant of the tethered DNA fragment and the persistence length can be derived from the two dimensional (2D) (x, y) projected image data. We have applied this method to various MgCl2 and glycerol concentrations as a proof of principle.

© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Heidelinde R. C. Dietrich ; Bernd Rieger ; Frank G. M. Wiertz ; Frederik H. De Groote ; Hendrik A. Heering, et al.
"Tethered particle motion mediated by scattering from gold nanoparticles and darkfield microscopy", J. Nanophoton. 3(1), 031795 (June 22, 2009). ; http://dx.doi.org/10.1117/1.3174445


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