In JoVE (1)
Other Publications (1)
Articles by Isabel Rathmann in JoVE
A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions Tanumoy Saha1, Isabel Rathmann1, Milos Galic1 1DFG Cluster of Excellence 'Cells in Motion', (EXC 1003), Institute of Medical Physics and Biophysics, University of Muenster We present a software solution for semi-automated tracking of relative protein concentration along the length of dynamic cellular protrusions.
Other articles by Isabel Rathmann on PubMed
Automated Analysis of Filopodial Length and Spatially Resolved Protein Concentration Via Adaptive Shape Tracking Molecular Biology of the Cell. Nov, 2016 | Pubmed ID: 27535428 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.