Note: Time-gated 3D single quantum dot tracking with simultaneous spinning disk imaging

Rev Sci Instrum. 2015 Dec;86(12):126102. doi: 10.1063/1.4937477.

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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Tracking / instrumentation*
  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement / instrumentation*
  • Imaging, Three-Dimensional / instrumentation*
  • Microscopy, Fluorescence / instrumentation*
  • Molecular Imaging / instrumentation*
  • Quantum Dots*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Signal Processing, Computer-Assisted / instrumentation