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MOUSE ORGAN HARVEST PROTOCOL 10/01 TO FREEZE TISSUES FOR FROZEN SECTIONS
 

Summary: MOUSE ORGAN HARVEST PROTOCOL 10/01
TO FREEZE TISSUES FOR FROZEN SECTIONS:
1. Make 1:1 OCT: PBS for tracheal infiltration to inflate lungs before freezing
2. Get bench area ready with absorbent material, styrofoam board and dissection equipment
3. In a small styrofoam flat container, make a slurry with small bits of dry ice and 2 methyl
butane. This will be the freezing mixture which will freeze the organs in OCT for frozen
sections
4. Lay euthanized animal on dissecting board
5. IF necessary, Perfuse animal with PBS and then with freshly made 4% paraformaldehyde
6. Expose trachea and infiltrate lungs with OCT/PBS mixture. Remove entire block and place on
paper towel to drain before freezing flat in vinyl mold with OCT.
7. Harvest tissues and place in sequence onto paper towels to dry off tissue fluids before placing into
vinyl molds, flat on the bottom and then fill with OCT. This keeps the tissues all at the same level:
A. spleen/thymus, lymph nodes. adrenal
B. Liver, pancreas, kidney
C. Heart, OCT inflated and drained lungs
D. GI (small intestine, colon, ? stomach) and GU (Bladder, prostate, testis, ovary, uterus)
E. Skin, skeletal muscle, salivary glands, mammary glands, (decalcified femur for blocks)
F. Brain
IT IS BETTER TO KEEP THESE AS SHOWN. CAN COMBINE A,B,C and D,E,F WHEN

  

Source: Abagyan, Ruben - School of Pharmacy and Pharmaceutical Sciences, University of California at San Diego

 

Collections: Biology and Medicine