Image artifacts in single molecule localization microscopy: why optimization of sample preparation protocols matters.

ABSTRACT: Single molecule localization microscopy (SMLM) techniques allow for sub-diffraction imaging with spatial resolutions better than 10 nm reported. Much has been discussed relating to different variations of SMLM and all-inclusive microscopes can now be purchased, removing the need for in-house software or hardware development. However, little discussion has occurred examining the reliability and quality of the images being produced, as well as the potential for overlooked preparative artifacts. As a result of the up to an order-of-magnitude improvement in spatial resolution, substantially more detail is observed, including changes in distribution and ultrastructure caused by the many steps required to fix, permeabilize, and stain a sample. Here we systematically investigate many of these ste

SUBMITTER: Whelan DR 

PROVIDER: S-EPMC4300460 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature