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J Vis Exp. 2014 Sep 7;(91):e52138. doi: 10.3791/52138.

Using plusTipTracker software to measure microtubule dynamics in Xenopus laevis growth cones.

Author information

1
Department of Biology, Boston College.
2
Department of Biology, Boston College; laura.lowery@bc.edu.

Abstract

Microtubule (MT) plus-end-tracking proteins (+TIPs) localize to the growing plus-ends of MTs and regulate MT dynamics(1,2). 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 polymerization(1). Many studies of EB1 behavior within growth cones have used time-consuming and biased computer-assisted, hand-tracking methods to analyze individual MTs(1-3). Our approach is to quantify global parameters of MT dynamics using the software package, plusTipTracker(4), following the acquisition of high-resolution, live images of tagged EB1 in cultured embryonic growth cones(5). 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 types(6-8).

PMID:
25225829
PMCID:
PMC4189079
DOI:
10.3791/52138
[Indexed for MEDLINE]
Free PMC Article

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