A single molecule investigation of the photostability of quantum dots

PLoS One. 2012;7(8):e44355. doi: 10.1371/journal.pone.0044355. Epub 2012 Aug 31.

Abstract

Quantum dots (QDs) are very attractive probes for multi-color fluorescence imaging in biological applications because of their immense brightness and reported extended photostability. We report here however that single QDs, suitable for biological applications, that are subject to continuous blue excitation from a conventional 100 W mercury arc lamp will undergo a continuous blue-switching of the emission wavelength eventually reaching a permanent dark, photobleached state. We further show that β-mercaptoethanol has a dual stabilizing effect on the fluorescence emission of QDs: 1) by increasing the frequency of time that a QD is in its fluorescent state, and 2) by decreasing the photobleaching rate. The observed QD color spectral switching is especially detrimental for multi-color single molecule applications, as we regularly observe spectral blue-shifts of 50 nm, or more even after only ten seconds of illumination. However, of significant importance for biological applications, we find that even small, biologically compatible, concentrations (25 µM) of β-mercaptoethanol has a significant stabilizing effect on the emission color of QDs, but that greater amounts are required to completely abolish the spectral blue shifting or to minimize the emission intermittency of QDs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescence
  • Photobleaching
  • Photochemical Processes*
  • Quantum Dots*
  • Spectrum Analysis

Grants and funding

This work was supported with grants from the Villum Kann Rasmussen Foundation to BioNET – Danish Center for Biophysics, and grants from Leo Pharma Forskningsfond, Lundbeckfonden and Novo Nordisk Fonden to BCL. This work was carried out in the laboratories of MEMPHYS – Center for Biomembrane Physics which is funded by the Danish Natural Research Foundation and using microscopy equipment at the Danish Molecular Biomedical Imaging Center (DaMBIC) which is funded by the Danish Ministry of Science. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.