Three-dimensional tracking of fluorescent nanoparticles with subnanometer precision by use of off-focus imaging.
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.
Speidel
Michael
M
Cell Biology and Biophysics Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Jonás
Alexandr
A
Florin
Ernst-Ludwig
EL
D005453:Fluorescence; D021621:Imaging, Three-Dimensional; D008962:Models, Theoretical; D036103:Nanotechnology; D055095:Optics and Photonics