A photoprotection strategy for microsecond-resolution single-molecule fluorescence spectroscopy.

Campos, Luis A; Liu, Jianwei; Wang, Xiang; Ramanathan, Ravishankar; English, Douglas S; Muñoz, Victor · Nat Methods · 2011

basic_science · Level V

Where this comes from

Abstract

Time resolution of current single-molecule fluorescence techniques is limited to milliseconds because of dye blinking and bleaching. Here we introduce a photoprotection strategy that affords microsecond resolution by combining efficient triplet quenching by oxygen and Trolox with minimized bleaching via the oxygen radical scavenger cysteamine. Using this approach we resolved the single-molecule microsecond conformational fluctuations of two proteins: the two-state folder α-spectrin SH3 domain and the ultrafast downhill folder BBL.

Medical subject headings