Unique in-house developed capabilities in active feedback for time-resolved 3D tracking of single nanoparticles, organic dyes, and fluorescent proteins.

Capabilities include:

• Explore 3D transport in live cells or selected soft materials.
• Can measure fluorescence lifetimes during tracking (3D tracking system).
• Can obtain wide-field contextual information from Nipkow Spinning Disk Microscopy.

Technical Specifications:

• For 2D tracking, 405, 488, 532, and 632 excitation wavelengths are possible.
• For 3D confocal tracking, the above wavelengths plus two-photon excitation at 880 nm also possible.
• 2D tracking performed at ~10 frames per second.
• 3D tracking stage position is updated 200 times per second.
• Accuracy depends upon tracking integration period — 3D tracking has ~30 nm (XY) and 100 nm (Z) accuracy for 5 millisecond integration period.

Contacts:
Jim Werner
Dean Morales

Research Highlights:
3D Particle Transport in Multichannel Microfluidic Networks with Rough Surfaces
Ryan, D. P.; Chen, Y.; Nguyen, P.; Goodwin, P. M.; Carey, J. W.; Kang, Q.; Werner, J. H.; Viswanathan, H. S. Scientific Reports 2020, 10 (1). doi.org/10.1038/s41598-020-70728-1

Three-Dimensional Single-Molecule Tracking in Living Cell
Kalb, D. M.; Ryan, D. P.; Morales, D. P.; Goodwin, P. M.; Werner, J. H. Spectroscopy and Dynamics of Single Molecules 2019, 229–267. doi.org/10.1016/B978-0-12-816463-1.00005-5

Multicolor Three-Dimensional Tracking for Single-Molecule Fluorence Resonance Energy Transfer Measurements
Keller, A. M.; DeVore, M. S.; Stich, D. G.; Vu, D. M.; Causgrove, T.; Werner, J. H. Analytical Chemistry 2018, 90 (10), 6109–6115. doi.org/10.1021/acs.analchem.8b00244