![]() ![]() Use the edge of the eyebrow hair to deepen the cleft formed by the involuting head mesoderm to allow peeling back to the Using an eyebrow knife, peel back the dorsal tissues to the dorsal blastopore lip ( Fig. Using one of the cutting methods described in Step 4, make a horizontal cut at the animal end of the dissection to completeĪ rectangle with the previous two incisions ( Fig. This trick requires some practice, but, once mastered, it is a clean and fast way to cut through thin tissues.ĥ. Alternatively, hold the region adjacent to the cutting edge with the hair loop, and gently flick upward with the eyebrow hair. This will complete the cut by pinching the tissue between the two hairs. Then press the hair loopĪgainst the tip of the eyebrow hair. Insert the tip of the eyebrow knife into one of the cuts, under the region that needs to be cut. If the cuts have not reached the animal pole, complete the cuts using one of the following approaches: Turn the embryo over, that is, animal pole facing up. Make a similar cut at the opposite end of the bottle cell line ( Fig. This cut is easily achieved by cutting against the base of the Petri dish, or by flicking the knife upward.Ĥ. Significant vertical juxtaposition of ectoderm and mesoderm has occurred. Explants are made at the onset of gastrulation before ![]() The ectoderm, mesoderm elongates in a plane with adjacent ectoderm. Resting on silicone vacuum grease until the desired stage, usually during or after neurulation. Explants are cultured beneath a coverslip fragment or a glass bridge With their inner surfaces apposed (closed sandwich). The explants are cultured flat, either as a single sheet (open-face explant) or more frequently as two sheets sandwiched together This is difficult to do when explants curl up, but in Keller sandwiches, Movements, particularly convergent extension, in culture. The purpose of Keller explants was initially to allow observation of gastrulation This protocol describes how to dissect,Īssemble, and cultivate Keller explants. It is ~60° to 90° wide, extending from the bottle cells to the animal pole. The basic Keller explant is a rectangle of dorsal mesendoderm and ectoderm from an early-gastrula-stage Xenopus laevis embryo. ![]()
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