Islet isolation (:

Hi! I'm back for episode 2!

Isolation of pancreatic islets from mice

Retrieved from BBC (2008). Scientists make weight loss claim. Retrieved on 01st July 2009, from: http://news.bbc.co.uk/2/hi/health/7372495.stm

For this experiment you will need these few things:
White mice, Anaesthesia, Surgical Scissors, surgical wire, Forceps, Syringe, Needle, Tube, Collagenase solution.

When I first stepped into the animal room, a strong smell engulfed me. At first I thought it smelt nice (maybe because it was before lunch), but as I babysit the mice longer, they started to stink. The mice were stored in a styrofoam with holes as they were transported from a facility (which I shall not name).

A total of 4 mice were used in this experiment. The mice were making a lot of noise and they kept moving around! They almost had the strength to push open the styrofoam cover and even tried to escape.

My senior picked up the mouse by its tail, and tried to hold up its head by its ears in between her thumb and index fingers and held the tail using her last finger. Basically, the mouse was immobilized by a hand. The other hand was used to inject the anaesthetic into the mouse’s abdomen. This has to be done carefully. If injected wrongly, this may cause the mice to die earlier, which will be no good for the experiment.

As you guys know, mouse is very small. Therefore my mentor used a binocular microscope for easier visualization. To ensure that the mouse is really asleep, she will pinch its legs to check for reaction. No reaction would indicate the mouse is asleep. Next, she used a surgical scissors to cut open the abdomen, and exposed its contents.

Do you know that a mouse’s liver is really big? At first, I thought the liver was the lung! Below the liver is the pancreas. First, my mentor skillfully tied up the bile duct with the surgical wire. Tying up the bile duct before injecting collagenase into the mice ensures that collagenase will only be injected in the pancreas. If not, collagenase solution will flow into the stomach and intestine, therefore the pancreas will not swell up. Next, she carefully inserted a small tube which contains collagenase into the bile duct. Collagenase is used to digest the external cells. Once collagenase is pumped into the pancreas, the pancreas starts to swell up. I once thought that the pancreas is green in colour, but actually it is quite yellowish-transparent. Once the entire amount is injected in, the tube was carefully removed and pancreas was removed and placed in a tube containing collagenase. The tube was then placed in 37*C waterbath. After the removal of pancreas, the mouse would die a few minutes later.

However, my observation did not stop here. As my senior needed to collect blood for her experiment, I watched as she inserted the needle into the mouse’s really small heart and took the small little amount of blood. It was really weird, as during the procedure, the mouse would have reflex action, almost like the mouse is having hiccups.

Lastly, after we have gotten what we needed, the mice were covered in aluminium foil for disposal.

after putting pancreas in collagenase, you'll incubate at 37oC for 17-18 minutes. after that, you'll shake the cells, to ensure homogenicity and check that there is no remnants. next, we will pour the solution into a petri dish. using a binocular microscope, islet cells are handpicked.the cells will be transferred into a petri dish containing RPMI (Roswell Park Memorial Institute) media which is suitable for islet growth. the cells. the handpicking of islet cells are repeated onto subsequent petri dishes to remove all the epithelial cells.
Although I wouldn't have a chance to ever cut the mouse open, but this has been a great experience, though this isn't and will not be my first and my last.

Lim Jia Hui (JOEY)
0703605F