title: Resolution doubling in 3D-STORM imaging through improved buffers.
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.
authors.affiliation: Laboratory for Experimental Biophysics, School of Basic Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland. nicolas.olivier@polytechnique.ed
authors.forename: Nicolas
authors.initials: N
authors.lastname: Olivier
authors.forename: Debora
authors.initials: D
authors.lastname: Keller
authors.forename: Pierre
authors.initials: P
authors.lastname: Gönczy
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence / Q000379:methods*; D011506:Proteins / Q000378:metabolism*
chemical_list: D011506:Proteins