Using plustiptracker software to measure microtubule dynamics in xenopus laevis growth cones

Alina Stout, Salvatore D'Amico, Tiffany Enzenbacher, Patrick Ebbert, Laura Anne Lowery

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Microtubule (MT) plus-end-tracking proteins (+TIPs) localize to the growing plus-ends of MTs and regulate MT dynamics1,2. One of the most well-known and widely-utilized +TIPs for analyzing MT dynamics is the End-Binding protein, EB1, which binds all growing MT plus-ends, and thus, is a marker for MT polymerization1. Many studies of EB1 behavior within growth cones have used time-consuming and biased computer-assisted, hand-tracking methods to analyze individual MTs1-3. Our approach is to quantify global parameters of MT dynamics using the software package, plusTipTracker4, following the acquisition of high-resolution, live images of tagged EB1 in cultured embryonic growth cones5. This software is a MATLAB-based, open-source, user-friendly package that combines automated detection, tracking, visualization, and analysis for movies of fluorescently-labeled +TIPs. Here, we present the protocol for using plusTipTracker for the analysis of fluorescently-labeled +TIP comets in cultured Xenopus laevis growth cones. However, this software can also be used to characterize MT dynamics in various cell types6-8.

Original languageEnglish
Article numbere52138
JournalJournal of Visualized Experiments
Issue number91
DOIs
StatePublished - 9 Jul 2014
Externally publishedYes

Keywords

  • EB1
  • Growth cone
  • Issue 91
  • Live cell imaging analysis
  • Microtubule dynamics
  • Microtubule plus-end-tracking proteins
  • Molecular Biology
  • PlusTipTracker
  • Xenopus laevis

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