Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy

Publikation: Beitrag in FachzeitschriftÜbersichtsartikel (Review)BeigetragenBegutachtung


  • Christopher Gell - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Volker Bormuth - (Autor:in)
  • Gary J Brouhard - (Autor:in)
  • Daniel N Cohen - (Autor:in)
  • Stefan Diez - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Claire T Friel - (Autor:in)
  • Jonne Helenius - (Autor:in)
  • Bert Nitzsche - , Core Facility Bedarfsorientierte Intelligente Mikroskopie (Autor:in)
  • Heike Petzold - (Autor:in)
  • Jan Ribbe - (Autor:in)
  • Erik Schäffer - , Nanomechanik (NFoG) (Autor:in)
  • Jeffrey H Stear - (Autor:in)
  • Anastasiya Trushko - (Autor:in)
  • Vladimir Varga - (Autor:in)
  • Per O Widlund - (Autor:in)
  • Marija Zanic - (Autor:in)
  • Jonathon Howard - (Autor:in)


In vitro assays that reconstitute the dynamic behavior of microtubules provide insight into the roles of microtubule-associated proteins (MAPs) in regulating the growth, shrinkage, and catastrophe of microtubules. The use of total internal reflection fluorescence microscopy with fluorescently labeled tubulin and MAPs has allowed us to study microtubule dynamics at the resolution of single molecules. In this chapter we present a practical overview of how these assays are performed in our laboratory: fluorescent labeling methods, strategies to prolong the time to photo-bleaching, preparation of stabilized microtubules, flow-cells, microtubule immobilization, and finally an overview of the workflow that we follow when performing the experiments. At all stages, we focus on practical tips and highlight potential stumbling blocks.


Seiten (von - bis)221-45
FachzeitschriftMethods in cell biology
PublikationsstatusVeröffentlicht - 2010

Externe IDs

Scopus 77955301639
ORCID /0000-0002-0750-8515/work/142235565



  • Animals, Cell Culture Techniques/methods, Cells, Cultured, Color, Fluorescent Dyes/pharmacology, Humans, Image Processing, Computer-Assisted/methods, Microscopy, Fluorescence/instrumentation, Microtubules/metabolism, Models, Biological, Staining and Labeling/methods, Tubulin/metabolism