Resolution doubling in 3D-STORM imaging through improved buffers.
Nicolas Olivier, Debora Keller, Pierre Gönczy, Suliana Manley
Index: PLoS ONE 8(7) , e69004, (2013)
Full Text: HTML
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.
Related Compounds
Related Articles:
2015-06-23
[Nucleic Acids Res. 43(11) , 5524-36, (2015)]
Identification of a novel self-sufficient styrene monooxygenase from Rhodococcus opacus 1CP.
2009-08-01
[J. Bacteriol. 191(15) , 4996-5009, (2009)]
Spatial Control of Epsin-induced Clathrin Assembly by Membrane Curvature.
2015-06-05
[J. Biol. Chem. 290 , 14267-76, (2015)]
2014-06-01
[Antimicrob. Agents Chemother. 58(6) , 3124-32, (2014)]
2013-04-01
[Indian J. Biochem. Biophys. 50(2) , 150-8, (2013)]