"Open-faced" explants are ones having the inner, deep cells exposed to the medium, often for the purpose of studying deep cell motility (39,46,66,67,70), and thus Danilchik's (DFA) medium is used. A culture chamber for these explants is made by drilling a 20-mm hole in a 60-mm plastic Petri dish and gluing a #1.5, 24-mm coverslip over the hole with silicone high vacuum grease (Dow Corning, Midland, MI). The tissue is placed with the inner or deep surface down on the coverslip, and restrained with another coverslip, supported at each end with silicone high vacuum grease. This allows high-resolution optics to be used with an inverted microscope. An upright microscope can be used by overfilling the dish with media, covering the top with a large glass slide, and then inverting it, such that the coverslip and the deep surface of the explant are at the top, facing the objective.
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