Group II caspase activity in bovine embryos【University of Florida】

2- Incubate the embryos in 25-ml microdrops of Hepes-TALP containing 5 mM PhiPhiLux G₁ D₂ at 39°C for 40 min in the dark.

3- Incubate negative controls in Hepes-TALP only. Following incubation, wash the embryos twice in 50-ml drops of Hepes-TALP

4- Place the embryos on poly-L-lysine coated slides (inside the thick circle made with the hydrophobic pen) and mount carefully with a cover slip so it does not crack the embryos.

5- Caspase activity must be determined immediately after the end of treatment (i.e., heat shock) using a Zeiss Axioplan 2 fluorescence microscope with a 453 objective.

Epifluoresence image of bovine embryos cultured at 38.5 or 41 C for 9 h