Resolving the orientation and position of single nanoparticles plays a significant role to understand the dynamics of the biological system. However, current full three-dimensional rotational tracking methods mainly rely on complex imaging pattern matching, which is time-consuming and easily disturbed by the complex cellular environments. Herein, we circumvent those problems by using dual-focus (both in-focus and defocused) dark field microscopy. By taking advantage of an auto feedback z adjustment system with the inclusion of a wedge prism in the light path, we could obtain the precious z position by keeping the gold nanorods on the perfect focal plane in one channel, while maintaining a consistent defocusing distance in the other channel. This new technique has allowed us to obtain valuable understandings on intracellular transport and endocytosis of nanoparticles and help develop an anti-cancer metastasis strategy with plasmonic gold nanoparticles.

Full 3D orientation and position determination of single anisotropic nanoparticles with dual-focus dark-field microscopy

256th National Meeting (2018)

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