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Tissue process for In Situ on paraffin embedded sections

*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 )

1. Fix tissue 4% Paraformadehyde for 48hours.
2. Wash with PBS 20 min X3 at 4°C.
3. Dehydrate:
   30% ETOH in ddH₂O for 1 hour at 4°C.
   50% ETOH in ddH₂O for 1 hour at 4°C.
   70% ETOH in ddH₂O for overnight or over weekend at 4°C .
   95% ETOH 3hour X 2 or overnight at 4°C.
   100% ETOH 1 hour X 2 at 4°C. Store at -20°C.
   100% ETOH 1 hour X 3 at RT.
4. Xylene 30 min X 2. Check embryos.
5. Paraffin 40 min X 3.
6. Embedding.
   
   (See more detail protocol at Histology protocols: How to fix and Dehydrate tissue)

 

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