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 paraffin 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:
- Euthanize the mouse by Isoflurane and neck-breaking.
- Use digital scale and weigh dish to make records of mouse body weight.
- Use pins to fix the mouse on a clean pad setting on a small foam board.
- Sanitize the mouse abdomen surface with 70% ethanol.
- Use scissors and tweezers to open the abdomen and dissect out the entire liver.
- Rinse liver in cold 1xPBS.
- Place the liver near a ruler with markings, take several photos.
- Make record of liver weight.
- Cut 2-3 small pieces of tissue from different liver lobes and put into Z-fix solution or 4% PFA (5ml/tube).
- 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).
- 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.
- Put the Eppendoff tubes and frozen blocks into -80°C.
- Put the tissue in Z-fix or 4% PFA tubes at 4°C.
- Put the dead bodies into a red bag and leave in -20°C.
- Transfer the photos from the camera to your own computer.
- Make record of the body weight and liver weight.
- The next morning, wash the fixed tissue with PBS and transfer it into cassettes. Soak the cassettes in 70% ethanol.