Single Coverslip with Liquid Medium OR Lying anf Hanging Drop Cultures, Harrison, Plasma, Explant, Coverslip, Embryo, Paraffin, Petroleum Jelly

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	Single Coverslip with Liquid Medium OR Lying anf Hanging Drop Cultures - This technique developed by Harrison (1907) has been most commonly used during the last more than fifty years and includes the following steps. (i) prepare medium in two parts, one containing 50% plasma in BSS (balanced salt solution) and the other containing 50% embryo extract in serum;
(ii) under sterile and aseptic conditions, using a capillary pipette, place one drop of plasma containing solution in the centre of each of one, or more coverslips (22mm); (iii) transfer a fragment (one or two pieces) of tissue (called explant) to this drop without crushing the tissue; (iv) add the embryo extract containing solution and mix thoroughly before clotting starts and then locate the explant; (v) place two small spots of petroleum jelly (using a glass rod) near the concavity of a depression slide and invert this slide over the coverslip; apply gentle pressure, so that jelly sticks to coverslip; (vi) allow culture medium to clot; (vii) turn over the slide and seal the margins of coverslip with paraffin; (viii) label and incubate at 37°C.