UCSF

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 Num

Manufacturer

1

ISOSOL (ISOFLURANE, USP)

 

Vedco Inc

2

Absorbent bench underpads (Trim to 1/4 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

Eppendoff 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 Eppendoff tube and then put the Eppendoff tube into a dry ice box immediately.
  12. Put the Eppendoff 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.