By David K. Gardner, Michelle Lane, Andrew J. Watson
This publication pulls jointly the entire diversity of cellphone tradition, biochemical, microscopic, and genetic suggestions to review the early mammalian embryo. in the past, there hasn't ever been this type of accomplished compendium, even though there were extra centred books of protocol, corresponding to Manipulating the Mouse Embryo, from chilly Spring Harbor. This ebook is meant to attract all constituencies, from uncomplicated experimental technological know-how to scientific and animal technology functions.
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Additional info for A Laboratory Guide to the Mammalian Embryo
5 Collection, Aspiration, Maturation, and Fertilization of Bovine Oocytes It is common to obtain slaughterhouse material for the generation of IVM/IVF/IVC (in vitro maturation/in vitro fertilization/in vitro culture) bovine embryos. 10. 6 Embryo Transfer in Mice In mice, embryos are transferred to pseudopregnant recipients. Pseudopregnant recipients are obtained by mating females to vasectomized males. Cleavage-stage embryos are usually transferred to the oviduct and blastocyst-stage embryos to the uterus.
To count the sperm for insemination, place 95 ml of water into a 5-ml tube. Using a different tip (as water will immobilize sperm), remove 5 ml of sperm and mix well with the water. Using a fresh tip, load 10 ml of this immobilized sperm into the top and bottom of the hemocytometer chamber. Count 5 squares (usually 4 corners and middle) of both the top and bottom. Average the counts. There should be between 30–40 sperm. This will result in an insemination volume of around 1 × 104 sperm/ml. If this is not the case, the insemination volume can be adjusted to compensate for low sperm numbers.
6. Cut the peritoneum to expose the body cavity. 7. Move the coils of gut to one side to expose the reproductive tract. 8. Hold the utero-tubal junction with watchmaker’s forceps, and, using the point of a pair of scissors, blunt dissect strip the surrounding connective tissue from the uterus. 9. For collection of the oviduct, make a cut between the oviduct and the ovary (avoid cutting the ovary). For collection of zygotes and two-cell embryos, make a second cut below the utero-tubal junction to isolate the oviduct from the uterus.