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PLoS One. 2013 Jul 17;8(7):e69004. doi: 10.1371/journal.pone.0069004. Print 2013.

Resolution doubling in 3D-STORM imaging through improved buffers.

Author information

1
Laboratory for Experimental Biophysics, School of Basic Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland. nicolas.olivier@polytechnique.ed

Abstract

Super-resolution imaging methods have revolutionized fluorescence microscopy by revealing the nanoscale organization of labeled proteins. In particular, single-molecule methods such as Stochastic Optical Reconstruction Microscopy (STORM) provide resolutions down to a few tens of nanometers by exploiting the cycling of dyes between fluorescent and non-fluorescent states to obtain a sparse population of emitters and precisely localizing them individually. This cycling of dyes is commonly induced by adding different chemicals, which are combined to create a STORM buffer. Despite their importance, the composition of these buffers has scarcely evolved since they were first introduced, fundamentally limiting what can be resolved with STORM. By identifying a new chemical suitable for STORM and optimizing the buffer composition for Alexa-647, we significantly increased the number of photons emitted per cycle by each dye, providing a simple means to enhance the resolution of STORM independently of the optical setup used. Using this buffer to perform 3D-STORM on biological samples, we obtained images with better than 10 nanometer lateral and 30 nanometer axial resolution.

PMID:
23874848
PMCID:
PMC3714239
DOI:
10.1371/journal.pone.0069004
[Indexed for MEDLINE]
Free PMC Article

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