Nonblinking and long-lasting single-molecule fluorescence imaging

Nat Methods. 2006 Nov;3(11):891-3. doi: 10.1038/nmeth934. Epub 2006 Oct 1.


Photobleaching and blinking of fluorophores pose fundamental limitations on the information content of single-molecule fluorescence measurements. Photoinduced blinking of Cy5 has hampered many previous investigations using this popular fluorophore. Here we show that Trolox in combination with the enzymatic oxygen-scavenging system eliminates Cy5 blinking, dramatically reduces photobleaching and improves the signal linearity at high excitation rates, significantly extending the applicability of single-molecule fluorescence techniques.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Carbocyanines / chemistry*
  • Chromans / chemistry*
  • Free Radical Scavengers / chemistry
  • Mercaptoethanol / chemistry*
  • Microscopy, Fluorescence / methods*
  • Oxygen / chemistry
  • Photobleaching
  • Sensitivity and Specificity
  • Time Factors


  • Carbocyanines
  • Chromans
  • Free Radical Scavengers
  • cyanine dye 5
  • Mercaptoethanol
  • 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
  • Oxygen