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

Philip Schäfer, Sebastian van de Linde, Julian Lehmann, Markus Sauer, Sören Doose

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

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.

LanguageEnglish
Pages3393-400
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number6
DOIs
Publication statusPublished - 19 Mar 2013

Fingerprint

Methylene Blue
Sulfhydryl Compounds
Oxygen
Cysteamine
Imaging techniques
Scavenging
Redox reactions
Fluorescence
Purging
Molecules
Molecular oxygen
Fluorescence spectroscopy
Bottles
Chromophores
Kinetic parameters
Microscopic examination
Kinetics
Enzymes
Experiments

Keywords

  • HeLa cells
  • spectrometry
  • methylene blue
  • microscopy, fluorescence/methods
  • microtubules/ultrastructure
  • fluorescence

Cite this

Schäfer, Philip ; van de Linde, Sebastian ; Lehmann, Julian ; Sauer, Markus ; Doose, Sören. / Methylene blue- and thiol-based oxygen depletion for super-resolution imaging. In: Analytical Chemistry. 2013 ; Vol. 85, No. 6. pp. 3393-400.
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Methylene blue- and thiol-based oxygen depletion for super-resolution imaging. / Schäfer, Philip; van de Linde, Sebastian; Lehmann, Julian; Sauer, Markus; Doose, Sören.

In: Analytical Chemistry, Vol. 85, No. 6, 19.03.2013, p. 3393-400.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Methylene blue- and thiol-based oxygen depletion for super-resolution imaging

AU - Schäfer, Philip

AU - van de Linde, Sebastian

AU - Lehmann, Julian

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