A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions

Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex signaling mechanisms underlying filopodial initiation, elongation and subsequent stabilization or retraction, it is crucial to determine the spatio...

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Bibliographic Details
Published in:Journal of visualized experiments 2017-07 (125)
Main Authors: Saha, Tanumoy, Rathmann, Isabel, Galic, Milos
Format: Article
Language:English
Subjects:
Animals
Cell Surface Extensions - chemistry
Cercopithecus aethiops
COS Cells
HeLa Cells
Humans
Molecular Biology
Pseudopodia - chemistry
Software - trends
Online Access:Get full text
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Description
Summary:Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex signaling mechanisms underlying filopodial initiation, elongation and subsequent stabilization or retraction, it is crucial to determine the spatio-temporal protein activity in these dynamic structures. To analyze protein function in filopodia, we recently developed a semi-automated tracking algorithm that adapts to filopodial shape-changes, thus allowing parallel analysis of protrusion dynamics and relative protein concentration along the whole filopodial length. Here, we present a detailed step-by-step protocol for optimized cell handling, image acquisition and software analysis. We further provide instructions for the use of optional features during image analysis and data representation, as well as troubleshooting guidelines for all critical steps along the way. Finally, we also include a comparison of the described image analysis software with other programs available for filopodia quantification. Together, the presented protocol provides a framework for accurate analysis of protein dynamics in filopodial protrusions using image analysis software.
ISSN:1940-087X
1940-087X
DOI:10.3791/55653