Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

This procedure begins by preparing a drosophila larva so that it has been turned inside out to expose the body wall muscle cell surface associated motor neurons and ggl cells by combining fluorescent staining for presynaptic neuron terminals, endogenous expression of GFP and DS red, targeted to glial and postsynaptic muscle cell membranes and visualization of the paras synaptic space Using fluorescent dyes, a clear picture of the structural changes occurring at the glial neuromuscular junction can be observed in a live preparation. Hi, I'm D Brink in Vanessa's ALS lab at the University of British Columbia in the zoology department. Today I'll show you a tissue preparation and two cell labeling techniques for imaging live al larvae tissue for confocal microscopy.

We use this procedure in the lab to study glial cells at the nerve muscle synapse. So now for something completely different, The inside out tissue preparation begins with staging of the larvae. This procedure will be demonstrated on wandering third larvae because they're big and easy to dissect, use only W three larvae that are actively crawling.