Automated analysis of filopodial length and spatially resolved protein concentration via adaptive shape tracking

Filopodia are dynamic, actin-rich structures that transiently form on a variety of cell types. To understand the underlying control mechanisms requires precise monitoring of localization and concentration of individual regulatory and structural proteins as filopodia elongate and subsequently retract...

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Bibliographic Details
Published in:Molecular biology of the cell 2016-11, Vol.27 (22), p.3616-3626
Main Authors: Saha, Tanumoy, Rathmann, Isabel, Viplav, Abhiyan, Panzade, Sadhana, Begemann, Isabell, Rasch, Christiane, Klingauf, Jürgen, Matis, Maja, Galic, Milos
Format: Article
Language:English
Subjects:
Actins - metabolism
Algorithms
Cell Shape - physiology
Computer Simulation
Image Processing, Computer-Assisted - methods
Pseudopodia - metabolism
Pseudopodia - physiology
Software
Spatio-Temporal Analysis
Statistics as Topic - methods
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Summary:Filopodia are dynamic, actin-rich structures that transiently form on a variety of cell types. To understand the underlying control mechanisms requires precise monitoring of localization and concentration of individual regulatory and structural proteins as filopodia elongate and subsequently retract. Although several methods exist that analyze changes in filopodial shape, a software solution to reliably correlate growth dynamics with spatially resolved protein concentration along the filopodium independent of bending, lateral shift, or tilting is missing. Here we introduce a novel approach based on the convex-hull algorithm for parallel analysis of growth dynamics and relative spatiotemporal protein concentration along flexible filopodial protrusions. Detailed in silico tests using various geometries confirm that our technique accurately tracks growth dynamics and relative protein concentration along the filopodial length for a broad range of signal distributions. To validate our technique in living cells, we measure filopodial dynamics and quantify spatiotemporal localization of filopodia-associated proteins during the filopodial extension-retraction cycle in a variety of cell types in vitro and in vivo. Together these results show that the technique is suitable for simultaneous analysis of growth dynamics and spatiotemporal protein enrichment along filopodia. To allow readily application by other laboratories, we share source code and instructions for software handling.
ISSN:1059-1524
1939-4586
DOI:10.1091/mbc.E16-06-0406