Harvest Liver Tissue from Mice

Bing Gui, Chen Lab, 5/31/2012

Prepare the tools and reagents needed for harvesting liver tissue from mice.

Items Catalog # Manufacturer
1 ISOSOL (ISOFLURANE, USP) Vedco Inc
2 Absorbent bench underpads (Trim to ¼ of its original size) 56636-032 VWR
3 70% ethanol Gold Shield Chemical Co.
4 Tweezer
5 Scissors
6 Cold 1x PBS UCSF CCF
7 Ruler
8 Digital Camera
9 Weigh Dish
10 Digital Scale
11 Cold 4% PFA or Z-fix (For parafin blocks)
12 Eppendorf tube (For frozen tissue)
13 Dry ice (For frozen blocks) UCSF CCF
14 2-Methylbutane (For frozen blocks) O35514 Fisher
15 Tissue-Tek® OCT™ Compound (For frozen blocks) SAK 4583 SAKURA
16 Red bag (For dead body)

Steps:

  1. Euthanize the mouse by Isoflurane and neck-breaking.
  2. Use digital scale and weigh dish to make records of mouse body weight.
  3. Use pins to fix the mouse on a clean pad setting on a small foam board.
  4. Sanitize the mouse abdomen surface with 70% ethanol.
  5. Use scissors and tweezers to open the abdomen and dissect out the entire liver.
  6. Rinse liver in cold 1xPBS.
  7. Place the liver near a ruler with markings, take several photos.
  8. Make record of liver weight.
  9. Cut 2-3 small pieces of tissue from different liver lobes and put into Z-fix solution or 4% PFA (5ml/tube).
  10. If O.C.T frozen blocks are required, cut 1-2 pieces of tissue and put it into a labeled mold with some O.C.T. Compound. After the tissue is placed on the mold, add some more O.C.T and then put the whole mold into 2-Methylbutane (chilling with dry ice).
  11. Cut the remained liver tissue into small pieces, put them into a labeled Eppendorf tube and then put the Eppendorf tube into a dry ice box immediately.
  12. Put the Eppendorf tubes and frozen blocks into -80°C.
  13. Put the tissue in Z-fix or 4% PFA tubes at 4°C.
  14. Put the dead bodies into a red bag and leave in -20°C.
  15. Transfer the photos from the camera to your own computer.
  16. Make record of the body weight and liver weight.
  17. The next morning, wash the fixed tissue with PBS and transfer it into cassettes. Soak the cassettes in 70% ethanol.