Stock Solutions

Pyruvate (20 mM) 22 mg / 10 ml saline. This can be stored frozen but not kept longer than 1 week.

Gentamycin 50 mg / ml in saline. Store at 5 C.

Acid Alcohol - Mix 1 part Glacial Acetic Acid with 3 part 95% Ethanol (e.g. 100 ml acetic acid and 300 ml ethanol).

Orecin Stain - 1% (weight/volume; 1 gm/100 ml); solvent is 60/40, water/acetic acid, filter berfore using.

PHE stock: (Penicillamine, Hypotaurine and Epinephrine) Hypotaurine-1mM, 1.09 mg/10 ml saline, aliquot and freeze Penicillamine-2mM, 3 mg/ 10ml saline, Aliquot and Freeze. Epinephrine- 250µM, Add 165 mg Na Lactate syrup (60%) to 50 mg Na metabisulfite to 50 ml ddH2O. Acidify to pH 4.0 with HCl. To 40 ml of this solution add 1.83 mg epinephrine. MAKE THIS STOCK JUST PRIOR TO MAKING PHE MIXTURE. For PHE Stock: 0.25 ml Penicillamine 0.25 ml Hypotaurine 0.10 ml Epinephrine 0.40 ml Saline Epinephrine is easily oxidized. Keep vials of PHE out of the light. Freeze and store. Thaw out of direct light and add 2 µl per 50 µl drop of fertilization media. Leibfried, M.L. and B.D. Bavister. 1982. J. Reprod. Fert. 66:87-93.

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