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Opt Lett. 2003 Jan 15;28(2):69-71.

Three-dimensional tracking of fluorescent nanoparticles with subnanometer precision by use of off-focus imaging.

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  • 1Cell Biology and Biophysics Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.


We show that the position of a fluorescent nanoparticle can be measured in three dimensions with subnanometer precision and 100-ms temporal resolution by use of standard epifluorescence video imaging in off-focus mode. The particle can be tracked without feedback in a volume of at least 40 microm x 60 microm x 3 microm. With the technique presented, the structure-mobility relationship of 216-nm latex particle in a porous polymer network was studied in three dimensions.

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