Methylene blue- and thiol-based oxygen depletion for super-resolution imaging.

Anal Chem 2013 Mar 1;85(6):3393-400. Epub 2013 Mar 1.

Department of Biotechnology & Biophysics, Biocenter, Julius-Maximilians-University Würzburg, Würzburg, Germany.

Anaerobic conditions are often required in solution-based bionanotechnological applications. Efficient oxygen depletion is essential for increasing photostability, optimizing fluorescence signals, and adjusting kinetics of fluorescence intermittency in single-molecule fluorescence spectroscopy/microscopy, particularly for super-resolution imaging techniques. We characterized methylene blue (MB)- and thiol-based redox reactions with the aim of designing an oxygen scavenger system as an alternative to the established enzyme-based oxygen scavenging systems or purging procedures. Redox reactions of the chromophore methylene blue in aqueous solution, commonly visualized in the blue bottle experiment, deplete molecular oxygen as long as a sacrificial reduction component is present in excess concentrations. We demonstrate that methylene blue in combination with reducing compounds such as β-mercaptoethylamine (MEA) can serve as fast and efficient oxygen scavenger. Efficient oxygen scavenging in aqueous solution is also possible with mere β-mercaptoethylamine at mM concentrations. We present kinetic parameters of the relevant reactions, pH-stability of the MB/MEA-oxygen scavenging system, and its application in single-molecule based super-resolution imaging.

Download full-text PDF

Source
http://pubs.acs.org/doi/10.1021/ac400035k
Publisher Site
http://dx.doi.org/10.1021/ac400035kDOI Listing
March 2013

Publication Analysis

Top Keywords

efficient oxygen
12
super-resolution imaging
12
methylene blue
12
redox reactions
8
aqueous solution
8
oxygen scavenger
8
oxygen depletion
8
oxygen scavenging
8
oxygen
7
blue bottle
4
bottle experiment
4
visualized blue
4
commonly visualized
4
methylene blue-
4
solution commonly
4
experiment deplete
4
sacrificial reduction
4
reduction component
4
long sacrificial
4
oxygen long
4

Similar Publications