Paraffin Embedding of Tissue Samples

From Bridges Lab Protocols
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Protocol

  1. After harvesting tissue from animal, place into pathology cassette and fix in 4% formaldehyde, 4% paraformaldehyde or 4% formalin O/N
  2. After fixation, begin dehydrating the tissue in 70% Ethanol
  3. On the day of planned embedding, wash sample with 100% Ethanol 3 times for 30 minutes.
    1. While washing in 100% Ethanol, start the paraffin wax machine (Link Building, Room 311)
  4. After 100% Ethanol washes, Wash 3 times in Xylenes
  5. Wash samples in a beaker filled with wax at 58-60C, 3 times for 15 mins
  6. Immediately before embedding, spray wax mold with "Mold Grease" and coat the bottom layer of the mold with wax
  7. Take your sample and orient it on top of the base wax as desired
  8. Place pathology cassette on top of the mold and fill with wax covering the mold
  9. Place mold with sample on "Cold Side" of embedding station
    1. Wax will solidify in 10-15 minutes
  10. Remove wax embedded sample from the mold (should just slip out)
  11. Store at room temperature until prepped to section