Filopodyan detects and accurately segments filopodia from a variety of different cell types in vitro and in vivo. (A) Growth cone from Xenopus retinal ganglion cell axons in culture, expressing GAP-RFP (a single time point from Video 1). (B) Tracheal cells in a Drosophila embryo expressing btl-Gal4 UAS-Cherry-CAAX (a single time point from Video 2). (C) Leading edge cells during dorsal closure in a Drosophila embryo expressing en-Gal4 UAS-cd8mCherry (a single time point from Video 3). (D) Human iPSC–derived cortical neurons in culture expressing cytoplasmic mNeonGreen (a single time point from Video 4). Bars, 5 µm.