1. Note: Time-gated 3D single quantum dot tracking with simultaneous spinning disk imaging
- Author
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Aaron M. Keller, James H. Werner, Mary E. Phipps, Matthew S. DeVore, Dominik G. Stich, Diane S. Lidke, Cédric Cleyrat, Jennifer A. Hollingsworth, Peter M. Goodwin, and Bridget S. Wilson
- Subjects
Microscope ,Photon ,Tracking (particle physics) ,Sensitivity and Specificity ,Fluorescence spectroscopy ,law.invention ,symbols.namesake ,Imaging, Three-Dimensional ,Optics ,law ,Notes ,Quantum Dots ,Microscopy ,Instrumentation ,Physics ,business.industry ,Reproducibility of Results ,Signal Processing, Computer-Assisted ,Equipment Design ,Image Enhancement ,Molecular Imaging ,Equipment Failure Analysis ,Microscopy, Fluorescence ,Cell Tracking ,symbols ,Time-resolved spectroscopy ,Molecular imaging ,business ,Raman scattering - Abstract
We describe recent upgrades to a 3D tracking microscope to include simultaneous Nipkow spinning disk imaging and time-gated single-particle tracking (SPT). Simultaneous 3D molecular tracking and spinning disk imaging enable the visualization of cellular structures and proteins around a given fluorescently labeled target molecule. The addition of photon time-gating to the SPT hardware improves signal to noise by discriminating against Raman scattering and short-lived fluorescence. In contrast to camera-based SPT, single-photon arrival times are recorded, enabling time-resolved spectroscopy (e.g., measurement of fluorescence lifetimes and photon correlations) to be performed during single molecule/particle tracking experiments.
- Published
- 2015