Now, I want to make paraffin wax with frozen tissue(Liver tissue). I check some information, it says that it is not suitable for frozen tissue to make paraffin wax, because tissue will be damaged during thawing.
Can someone give me some suggestion about this? What is the best way to make paraffin wax with frozen tissue?
Thank you for your answer.
Dachen Zhou
Hi Dachen, it depends weather your tissue is fixed or not. Take a piece of liver for testing. If it is formalin fixed let it thaw in PBS in the fridge, change the buffer several times; after the tissue is thawn start the embedding with ascended alcohols for dehydration, Xylol and Paraffin. If your tissue is unfixed thaw it carefully from -80°C to -20°C to Formalin or 4% PFA in the fridge. lieve it there for 2-3 days (depends on the sice of the tissue), rinse in PBS and start with the embedding protocol. Take sections from these blocks and make a H&E stain to check the quality of the cell structure. If it is ok continue, if not take frozen sections from the cryostate. Good luck!
Hi Dachen, it depends weather your tissue is fixed or not. Take a piece of liver for testing. If it is formalin fixed let it thaw in PBS in the fridge, change the buffer several times; after the tissue is thawn start the embedding with ascended alcohols for dehydration, Xylol and Paraffin. If your tissue is unfixed thaw it carefully from -80°C to -20°C to Formalin or 4% PFA in the fridge. lieve it there for 2-3 days (depends on the sice of the tissue), rinse in PBS and start with the embedding protocol. Take sections from these blocks and make a H&E stain to check the quality of the cell structure. If it is ok continue, if not take frozen sections from the cryostate. Good luck!
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