Biological Insight from Super-Resolution Microscopy: What We Can Learn from Localization-Based Images

Annu Rev Biochem. 2018 Jun 20:87:965-989. doi: 10.1146/annurev-biochem-060815-014801. Epub 2017 Dec 22.

Abstract

Super-resolution optical imaging based on the switching and localization of individual fluorescent molecules [photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), etc.] has evolved remarkably over the last decade. Originally driven by pushing technological limits, it has become a tool of biological discovery. The initial demand for impressive pictures showing well-studied biological structures has been replaced by a need for quantitative, reliable data providing dependable evidence for specific unresolved biological hypotheses. In this review, we highlight applications that showcase this development, identify the features that led to their success, and discuss remaining challenges and difficulties. In this context, we consider the complex topic of defining resolution for this imaging modality and address some of the more common analytical methods used with this data.

Keywords: PAINT; PALM; STORM; nanoscopy; quantitative analysis; single-molecule localization microscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Algorithms
  • Animals
  • Cluster Analysis
  • Fourier Analysis
  • Humans
  • Imaging, Three-Dimensional
  • Models, Biological
  • Molecular Structure
  • Nanotechnology
  • Single Molecule Imaging / methods*
  • Single Molecule Imaging / statistics & numerical data
  • Stochastic Processes