Review: dSTORM - Single-molecule photoswitching and localization

05/30/2011

S. van de Linde, S. Wolter, M. Sauer. Aus. J. Chem. 64, 503-511 (2011)

Super-resolution imaging methods based on single-molecule localization are reviewed. The use of such methods to successfully study various cellular structures in fixed and living cells with quite recently unforeseeable optical resolution is demonstrated. Applications to three-dimensional imaging are also highlighted.
Super-resolution imaging methods based on single-molecule localization are reviewed. The use of such methods to successfully study various cellular structures in fixed and living cells with quite recently unforeseeable optical resolution is demonstrated. Applications to three-dimensional imaging are also highlighted.

Abstract

Within only a few years super-resolution fluorescence imaging based on single-molecule localization and image reconstruction has attracted considerable interest because it offers a comparatively simple way to achieve a substantially improved optical resolution down to ~20 nm in the image plane. Since super-resolution imaging methods such as photoactivated localization microscopy, fluorescence photoactivation localization microscopy, stochastic optical reconstruction microscopy, and direct stochastic optical reconstruction microscopy rely critically on exact fitting of the centre of mass and the shape of the point-spread-function of isolated emitters unaffected by neighbouring fluorophores, controlled photoswitching or photoactivation of fluorophores is the key parameter for resolution improvement. This review will explain the principles and requirements of single-molecule based localization microscopy, and compare different super-resolution imaging concepts and highlight their strengths and limitations with respect to applications in fixed and living cells with high spatio-temporal resolution.

Australian Journal of Chemistry

Back