A simple, versatile method for GFP-based super-resolution microscopy via nanobodies

Nat Methods. 2012 Jun;9(6):582-4. doi: 10.1038/nmeth.1991. Epub 2012 Apr 29.


We developed a method to use any GFP-tagged construct in single-molecule super-resolution microscopy. By targeting GFP with small, high-affinity antibodies coupled to organic dyes, we achieved nanometer spatial resolution and minimal linkage error when analyzing microtubules, living neurons and yeast cells. We show that in combination with libraries encoding GFP-tagged proteins, virtually any known protein can immediately be used in super-resolution microscopy and that simplified labeling schemes allow high-throughput super-resolution imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fluorescent Dyes
  • GPI-Linked Proteins / immunology*
  • Green Fluorescent Proteins / immunology*
  • Microscopy, Fluorescence / methods*
  • Nanotechnology
  • Neurons / ultrastructure
  • Rats
  • Saccharomycetales / ultrastructure


  • Fluorescent Dyes
  • GPI-Linked Proteins
  • Green Fluorescent Proteins