TY - JOUR
T1 - Using plustiptracker software to measure microtubule dynamics in xenopus laevis growth cones
AU - Stout, Alina
AU - D'Amico, Salvatore
AU - Enzenbacher, Tiffany
AU - Ebbert, Patrick
AU - Lowery, Laura Anne
N1 - Publisher Copyright:
© JoVE 2006-2014. All Rights Reserved.
PY - 2014/7/9
Y1 - 2014/7/9
N2 - 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.
AB - 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.
KW - EB1
KW - Growth cone
KW - Issue 91
KW - Live cell imaging analysis
KW - Microtubule dynamics
KW - Microtubule plus-end-tracking proteins
KW - Molecular Biology
KW - PlusTipTracker
KW - Xenopus laevis
UR - http://www.scopus.com/inward/record.url?scp=84908274986&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84908274986&partnerID=8YFLogxK
U2 - 10.3791/52138
DO - 10.3791/52138
M3 - Article
C2 - 25225829
AN - SCOPUS:84908274986
SN - 1940-087X
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 91
M1 - e52138
ER -