Tsunoyama, T. A. et al. Nat. Chem. Biol. 14, 497–506 (2018).

Single-molecule tracking can provide important insight into protein dynamics, regulation and function. However, photophysical properties of fluorophores, such as bleaching and blinking, can make it challenging to track labeled molecules over extended periods. Although methods for reducing photobleaching and photoblinking are known, they can be incompatible with live imaging because of their toxicity. Tsunoyama et al. addressed this challenge by treating cells with low concentrations of dissolved oxygen along with a reducing-plus-oxidizing system to suppress both bleaching and blinking. They demonstrate that the approach works for numerous commonly used organic dyes and has only minor effects on cells. Using their approach, they extended the duration of particle tracking from ~10 seconds to ~7 minutes at video rate, while maintaining high localization precision. They used their approach to study the behavior of integrins at focal adhesions.