实验动物基本参数

ABSTRACT Knowledge of the geneological relationships of cells during development can allow one to gain insight into when and where developmental decisions are being made. Geneological relationships ca ...

Procedures for In Vitro Production of Bovine Embryos The procedures for in vitro production in vitro fertilization (IVF) and in vitro culture (IVC) of embryos described here are based on procedures d ...

8 eggs per day day 7- day 13 cut CAM wash in precooled PBS in 10 ml WASH 1 (PBS 5 mM EDTA COMPLETE) on ice cut in pieces in petri dish on ice centrifuge 4 min at 1000 rpm (table top centifuge Falcon 1 ...

1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forcepsuse another to tear the mesometrium away from the uterus.3. Remove the embryos fr ...

1. The basic unit is constructed from a media filtration unit such as Corning’s 115 ml filter system (catalog numbers 430944-430947). 2. Tygon tubing (3.2 mm i.d. x 6.4 mm o.d.) with a stopcock fasten ...

MEMFA Fix10xMEMFA Salts1 part 10x MEMFA salts1 M MOPS1 part 37% formaldehyde20mM EGTA8 parts water10mM MgSO410x salts can be autoclaved and stored. Turns yellow after autoclaving. ...

Ovulation is induced in X. tropicalis by injection of human chorionic gonadotropin (hCG). X. tropicalis requires a much smaller dose than does X. laevis. We use basically the same protocol as that des ...

When we first started using X. tropicalis in vitro fertilization had an extremely poor efficiency. However with the careful selection of a mature male and the use of L15/CS (L15 Leibovitch's Media sup ...

In order to get the best in vitro fertilization two criteria appear to be critical: first selection of a mature male and second use of L15/CS (L-15 Medium supplemented to 10% calf serum).Select a mal ...

For natural matings single pairs of boosted males and females are placed together. We usually keep them on a darkened shelf away from disturbances but where they can be easily monitored. Three to four ...

Four hours after boosting with hCG the females are ready to begin laying eggs. The best test is to see if there are eggs in the holding buckets. Also the cloaca gets red and swollen like in X. laevis. ...

Spawning can be induced in adult sea urchinsby injecting 1 ml of a0.5M KCl solution into several sites in the soft membrane around the mouth. Within minutes the gametes should appear: the sperm are of ...

Gametes are collected as described. To remove some of their jelly coatwash eggs several times in ASW by allowing them to settle and gently pouring off excess liquid. Transfer enough eggs to barely cov ...

1. Transfer 50 ml of embryo cultures to centrifuge tubes. Spin at 1500 rpm for 5 minutes. Check that you can see a pellet of embryos at the bottom.Quickly pour off the ASW. Try to remove as much as po ...

Histochemical staining of sea urchin embryos for alkaline phosphatase (AP) enzyme activity1. Obtain embryo samples tube of AP substrate buffer and tube of phosphate buffered saline (PBS) for each grou ...

1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 100 ul 1% normal goat serum to the other as a contr ...

The following is written to help non-marine biologists use sea urchins occasionally in teaching; disregard if you have running sea water tanks in your lab!Gravid animals can be obtained virtually year ...

Solutions for Sea UrchinASW: Artificial sea water (MBL)NaCl24.72 gKCl0.67 gCaCl2.2H2O1.36 gMgCl2.6H2O4.66 gMgSO4.7H2O6.29 gNaHCO30.18 gdH2Oto 1 literSulfate-free Sea waterNaCl27.72 gKCl0.67 gCaCl2.2H2 ...

Invertebrate embryos develop at different rates depending on the temperature of incubation. Descriptive stages based on the number of cells and their arrangement in the embryo are used to enable resea ...

Jill K. Flemming Franklin & Marshall College Class of 2001 IntroductionThe objective of this project is to observe the effects of UV radiation on both fertilization and development in the sea urchin e ...