easySTORM: a robust, lower-cost approach to localisation and TIRF microscopy

J Biophotonics. 2016 Sep;9(9):948-57. doi: 10.1002/jbio.201500324. Epub 2016 Mar 11.

Abstract

TIRF and STORM microscopy are super-resolving fluorescence imaging modalities for which current implementations on standard microscopes can present significant complexity and cost. We present a straightforward and low-cost approach to implement STORM and TIRF taking advantage of multimode optical fibres and multimode diode lasers to provide the required excitation light. Combined with open source software and relatively simple protocols to prepare samples for STORM, including the use of Vectashield for non-TIRF imaging, this approach enables TIRF and STORM imaging of cells labelled with appropriate dyes or expressing suitable fluorescent proteins to become widely accessible at low cost.

Keywords: Microscopy; PALM; STORM; localization; super-resolution.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Endothelial Cells / cytology
  • Fibroblasts / cytology
  • Fluorescent Dyes
  • Humans
  • Lasers
  • Light
  • Mice
  • Microscopy, Fluorescence / economics
  • Microscopy, Fluorescence / methods*
  • Optical Fibers
  • Proteins
  • Software

Substances

  • Fluorescent Dyes
  • Proteins