Just before use (i.e., ~10 min) prepare the following in a 15
mL conical tube covered with aluminum foil:
100 μl EtBr Stock (0.05 mg/mL)
3 μl FDA Stock (0.005 mg/mL)
10 mL DPBS or culture medium
Note: If you want to recover embryos from glass slide, use DPBS with 0.1% BSA or serum to prevent them from sticking to slide.
3) Place on ice until needed.
4) To stain embryos or oocytes place 50 μl of dye solution on a glass slide.
5) In the smallest volume possible transfer embryos or oocytes to be stained in the 50 μl and allow to sit in the dark for at least 3 min (FDA cleavage of acetate radical traps dye inside cell; 3 min=time for accumulation).
6) View embryos or oocytes for staining using the fluorescence microscope under UV epiluminesence (use UV filter).
Live Stain=Green
Dead Stain=Red/Orange
Count green first before it "burns out" from illumination
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