Computational protocol: WASP-Arp2/3-dependent actin polymerization influences fusogen localization during cell-cell fusion in Caenorhabditiselegans embryos

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Protocol publication

[…] C. elegans eggs in M9 buffer were mounted on 3% agarose pads at 20°C (). Live-cell images were collected with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging, Jena, Germany) equipped with a 100×/1.45 NA objective or a IX83 microscope (Olympus, Southend-on-Sea, UK) equipped with a 150×/1.45 NA oil objective, an EM CCD camera (Andor iXon+DU-897D-C00-#BV-500, Andor Technology, Belfast, UK), and the 488 nm and 568 nm lines of a Sapphire CW CDRH USB Laser System (Coherent, Santa Clara, USA) with a spinning disk confocal scan head (CSU-X1 Spinning Disk Unit, Yokogawa, Kanazawa, Japan). Time-lapse images were acquired with an exposure time of 200 ms every 2 min for imaging the entire cell-cell fusion process. Images were acquired with μManager software (https://www.micro-manager.org/), and processed and quantified with ImageJ software (http://rsbweb.nih.gov/ij/). […]

Pipeline specifications

Software tools μManager, ImageJ
Applications SPIM, Microscopic phenotype analysis
Organisms Drosophila melanogaster, Caenorhabditis elegans