Deoxyglucose Uptake Assay

1. Blastocysts are moved by mouth pipet to HTF media supplemented with D-glucose to give a final concentration of 5.6 mM and containing 500 nM insulin for 30 min. Following this preincubation, they are moved in groups of 10-20 into Ringers wash and then into media containing 200 iM 2-DG for 15 min. A basal group is placed in HTF with 5.6 mM D-glucose with no insulin added.

2. The embryos are then removed, washed in DG-free, BSA-free Ringers solution for 1 min, and then transferred with 0.5-1 ^L of the same salt solution onto a glass slide with a braking pipet. The embryo is then quick-frozen immediately by dipping the glass slide into isopentane brought to its freezing point (— 170°C) with liquid N2.

3. The specimens are then freeze-dried on the slide at — 35°C in a glass vacuum tube at a vapor pressure of <0.01 mm Hg. The slides are then stored at —20°C under reduced pressure.

Supplements For Diabetics

Supplements For Diabetics

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