(A) We label retinal ganglion cell growth (RGC) growth cones in live embryos at approximately 30 hours post-fertilization by placing them in a solution of methylcellulose. We then inject the fluorescent, lipophilic dye, DiI, into one eye and allow the dye to diffuse. (B) After a few hours, we embed several embryos in a droplet of agarose made up in embryo medium containing tricaine and image using a laser scanning confocal microscope. (C) A series of optical sections through the growth cone and its trailing axon are imaged every 3 minutes. The z-series at each time point is projected into two dimensions and the resulting images are combined into time-lapse movies such as those seen below. (Thanks to Charles Holly Jr. for writing the java plugin enabling composition of movies.)