Thursday, October 25, 2012

Well, we finally got started running our experiments today. The first thing I  had to do was to make sure that I had everything that I needed, ready to go in the cold room. The cold room is at 0 degrees Celsius (32 F). We need keep it that cold because the fish live at -1.86 C (28.7 F) and they get very stressed out at even +2 C. However, a lot of our instruments etc don't function below 0 C so that is the compromise we need to make. Here is our lab bench in the cold room.

Next, I made sure that all of our water baths were at the correct temperatures for our experiments. We will be conducting our heat shock experiments using a range of temperatures: -2 C (which is our control), 0 C, +2 C, +6 C (a lethal temperature for most of the fish we are studying), and +10 C. Here are what our water baths look like.

Next we needed a "volunteer." This isn't the actual fish we used today, but it is a picture of what a Trematomus bernacchii looks like.

After I sacrificed the fish, I removed its liver and perfused (pumped out) the blood from it as well. Then I sectioned it into small pieces before placing it into a bath of collagenase which helps to break up the tissue into individual cells. Here is some liver tissue floating in collagenase.

Next, I had to crush the liver tissue into very small (microscopic) pieces to isolate the individual cells. Here I am doing that!! Remember, it is 0 degrees C (28.7 F) in the cold room. Working in there for extended periods of time gets pretty cold. And there are fans blowing the cold air around too. Big Red is a must for this stage of my lab work!

And here is my "slurry" of liver cells (called hepatocytes)!

For those of you who followed my blog last year, this next part will look very familiar! Tubes floating in water! Each water bath is set at a different temp. The tubes have a special solution in them that is identical to the fish's blood (without blood cells) so that they can continue to function long after they have been removed from the fish. Last year, I made little foam square and poked holes into them to make the perfect kind of floatation devices for my tubes. I was so happy to see that they were still here in Crary when we got back this year!

It felt so good to finally start some lab work. Don't get me wrong, I love fishing! But that is only one very small part of what we are here to do and I was excited to get the next stage of our research started. Everything ran smoothly, which made for another great day!