TY - JOUR N1 - © 2017 Urban?i? et al. https://creativecommons.org/licenses/by/4.0/ This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). IS - 10 SP - 3405 VL - 216 JF - Journal of Cell Biology A1 - Urban?i?, V A1 - Butler, R A1 - Richier, B A1 - Peter, M A1 - Mason, J A1 - Livesey, FJ A1 - Holt, CE A1 - Gallop, JL SN - 1540-8140 UR - http://dx.doi.org/10.1083/jcb.201705113 TI - Filopodyan: An open-source pipeline for the analysis of filopodia EP - 3422 AV - public Y1 - 2017/10/02/ KW - Animals KW - Cell Line KW - Drosophila melanogaster KW - Embryo KW - Nonmammalian KW - Humans KW - Image Processing KW - Computer-Assisted KW - Microscopy KW - Fluorescence KW - Pseudopodia KW - User-Computer Interface KW - Xenopus laevis ID - discovery10056832 N2 - Filopodia have important sensory and mechanical roles in motile cells. The recruitment of actin regulators, such as ENA/VASP proteins, to sites of protrusion underlies diverse molecular mechanisms of filopodia formation and extension. We developed Filopodyan (filopodia dynamics analysis) in Fiji and R to measure fluorescence in filopodia and at their tips and bases concurrently with their morphological and dynamic properties. Filopodyan supports high-throughput phenotype characterization as well as detailed interactive editing of filopodia reconstructions through an intuitive graphical user interface. Our highly customizable pipeline is widely applicable, capable of detecting filopodia in four different cell types in vitro and in vivo. We use Filopodyan to quantify the recruitment of ENA and VASP preceding filopodia formation in neuronal growth cones, and uncover a molecular heterogeneity whereby different filopodia display markedly different responses to changes in the accumulation of ENA and VASP fluorescence in their tips over time. ER -