Focus on Super-Resolution Imaging with Direct Stochastic Optical Reconstruction Microscopy (dSTORM)
Donna R. Whelan A , Thorge Holm B , Markus Sauer B and Toby D. M. Bell A CA School of Chemistry, Monash University, Wellington Road, Clayton, Vic. 3800, Australia.
B Julius-Maximilians University of Würzburg, Am Hubland, 97074 Würzburg, Germany.
C Corresponding author. Email: toby.bell@monash.edu
Australian Journal of Chemistry 67(2) 179-183 https://doi.org/10.1071/CH13499
Submitted: 19 September 2013 Accepted: 20 November 2013 Published: 13 December 2013
Abstract
The last decade has seen the development of several microscopic techniques capable of achieving spatial resolutions that are well below the diffraction limit of light. These techniques, collectively referred to as ‘super-resolution’ microscopy, are now finding wide use, particularly in cell biology, routinely generating fluorescence images with resolutions in the order of tens of nanometres. In this highlight, we focus on direct Stochastic Optical Reconstruction Microscopy or dSTORM, one of the localisation super-resolution fluorescence microscopy techniques that are founded on the detection of fluorescence emissions from single molecules. We detail how, with minimal assemblage, a highly functional and versatile dSTORM set-up can be built from ‘off-the-shelf’ components at quite a modest budget, especially when compared with the current cost of commercial systems. We also present some typical super-resolution images of microtubules and actin filaments within cells and discuss sample preparation and labelling methods.
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