Nonblinking and long-lasting single-molecule fluorescence imaging.
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.
Rasnik
Ivan
I
Department of Physics, Emory University, Atlanta, Georgia 30322, USA. irasnik@physics.emory.edu
McKinney
Sean A
SA
Ha
Taekjip
T
D002232:Carbocyanines / Q000737:chemistry*; D002839:Chromans / Q000737:chemistry*; D016166:Free Radical Scavengers / Q000737:chemistry; D008623:Mercaptoethanol / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010100:Oxygen / Q000737:chemistry; D038761:Photobleaching; D012680:Sensitivity and Specificity; D013997:Time Factors
D002232:Carbocyanines; D002839:Chromans; D016166:Free Radical Scavengers; C085321:cyanine dye 5; D008623:Mercaptoethanol; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D010100:Oxygen