Tissue Process for Whole Mount In Situ
*The following wash and dehydration steps should be carried out at 4 °C on a shaker.( No shaking with Day 8.5-9.5 embryos )
*Use all RNase free solutions.
- Fix embryos overnight in 4% paraformaldehyde/PBS at 4°C with shaking.
- Wash with 1min PBT(PBS+0.1%Tween-20).
- Wash 3X 10 min with PBT at 4°C.
- Dehydrate :
25% methanol in PBT 20-40min.
50% methanol in PBT 20-40min.
75% methanol in PBT 20-40min.
100% methanol in PBT 20-40min.
Change to new methanol.
- Store at -20°C until further use.