Single-molecule imaging of non-equilibrium molecular ensembles on the millisecond timescale

Nat Methods. 2016 Apr;13(4):341-4. doi: 10.1038/nmeth.3769. Epub 2016 Feb 15.


Single-molecule fluorescence microscopy is uniquely suited for detecting transient molecular recognition events, yet achieving the time resolution and statistics needed to realize this potential has proven challenging. Here we present a single-molecule imaging and analysis platform using scientific complementary metal-oxide semiconductor (sCMOS) detectors that enables imaging of 15,000 individual molecules simultaneously at millisecond rates. This system enabled the detection of previously obscured processes relevant to the fidelity mechanism in protein synthesis.

Publication types

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

MeSH terms

  • Algorithms
  • Bacteria / ultrastructure
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods*
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods*
  • RNA, Transfer / ultrastructure*
  • Ribosomes / ultrastructure*
  • Time Factors


  • RNA, Transfer