title: On the mechanism of Trolox as antiblinking and antibleaching reagent.
abstract: Recent advances in photobleaching and blinking prevention have aided the advancement of single-molecule and super-resolution fluorescence microscopy. However, a common mechanism of the action of antifading agents such as Trolox is still missing. In this communication we present evidence that Trolox acts in accordance with a mechanism that involves triplet quenching through electron transfer and subsequent recovery of the resulting radical ion by the complementary redox reaction. The required oxidant for this unifying mechanism based on a reducing and oxidizing system (ROXS) is formed via (photo-) reaction with molecular oxygen. We present evidence that this oxidized form is a quinone derivative of Trolox with strong oxidizing properties. These findings shed light on many contradicting results regarding the action of antifading agents and might lead to a common mechanistic understanding of photobleaching and its prevention. Finally, a recipe on the proper use of Trolox as an antifading agent is provided.
authors.affiliation: Angewandte Physik-Biophysik, and Center for NanoScience, Ludwig-Maximilians-Universitat, Amalienstrasse 54, 80799 Munchen, Germany.
authors.forename: Thorben
authors.initials: T
authors.lastname: Cordes
authors.forename: Jan
authors.initials: J
authors.lastname: Vogelsang
authors.forename: Philip
authors.initials: P
authors.lastname: Tinnefeld
mesh_terms: D016227:Benzoquinones / Q000737:chemistry; D002839:Chromans / Q000737:chemistry*; D004579:Electron Transport; D005456:Fluorescent Dyes / Q000737:chemistry*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry*; D038761:Photobleaching
chemical_list: D016227:Benzoquinones; D002839:Chromans; D005456:Fluorescent Dyes; C004532:quinone; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D010100:Oxygen