Pieces of Pullman (part I) : winco

So my first real blog about nothing associated with school, but about life in Pullman in general. Or in this case, Moscow, ID, which is where my supermarket, Winco, is located. It's definitely a far cry from the Trader Joe's and Whole Foods that I've left behind, and more of a cross between Costco (price-wise) and Super A (atmosphere-wise). ...I know. you can't believe I'm shopping here.

<--But check this out! ALL THAT FOOD!!! FOR $44.09!!! 2 fillets of snapper, buttersquash soup, tomato soup, milk, wheat bread, udon, yogurt, veggies, the list goes on. It blows my mind. Another idea I'm completely drawn to is their cereal aisle. They have the normal boxes too, but this way, I can have a different bowl of cereal everyday for 30 days! How amazing! Theoretically I could just buy 7 different boxes of cereal and alternate between boxes, but I'm one of those people who feels guilty about opening a new box before the old one is finished. Then there are those little tiny-individual-bowl boxes of cereal, but that option consists of way too much packaging and is consumerism to the max. This way, I have my cereal variety and my mind is at ease. The barrels on the floor are different baking mixes-- muffin mix, cornbread mix, chocolate cake mix, etc. so you don't need to buy the boxes and you can buy as much or as little as you want.


And, as my mom pointed out when she was here, they have some of the asian stuff I wouldn't have expected them to. i.e wonton skins, BOTH types of tofu, and more than one kind of soy sauce. The Japanese student in my lab, So, told me the quality of fish here is not very good, but considering I can't go 6 months without fish, I've made do. So far I've tried the sockeye salmon, red snapper, and dover sole. The dover sole has been the best was the best so far. I've got a catfish fillet still in the fridge, but my hopes aren't high. They are all however, wild-caught and pretty fresh by the packaging date. I'll lay off the red snapper since I realized its an eco-worst fish. Sad, since my recipe was really delicious.

egg sucking

It is no longer the hot summer it once was. The days so far have been beautiful. The campus is speckled with these trees--half orange and red, the other half still green and yellow, that show a rainbow of autumn colors. Again, its just been gorgeous. The fields, however, are a little bare since the harvest has come and gone. Soon though there will be new crop.

School has been fine; 2 weeks til first exams. Rotation has been going well, concentrating on improving my egg sucking skills. Getting better at identifying them, but still moving slower than I should. A brief explanation into the technical work of egg sucking:

What you are looking for is the germinal vesicle oocytes (Figure 1). This is a clean picture. In reality, you must hunt for these in a brownish pink solution amongst bad oocytes and secondary (larger) oocytes you don't want, and all other cell types out of the ovary that you just mascerated.

Figure 1. Notice that clear circle in the middle of the egg. Within that, is the nucleus. We refer to these eggs as the ones with the 'winkies'.

A good micropipette can make or break you.
1) Pipette is too big-- this will just suck up everything.
2) Pipette is too small--can dent your oocytes and also becomes easily clogged with other unwanted cells.
It will take you longer if you encounter these issues, and oocytes are very fickle, they start degenerating if you don't move quickly. They start looking like this (Figure 2) and even this (Figure 3).

Figure 2. The sides of the oocytes become dented and the nucleus is no longer visible.

Figure 3. The zona pellucida surrounding the oocyte has become thick and dark granules appear inside the oocyte.

The oocytes should not really sit for more than 15 minutes. After collection, you culture them in an incubator overnight, then check how many of them have produced a polar body (Figure 4). Collecting them and putting them in culture inspires the oocytes to undergo meiosis I, to produce the polar body.

Figure 4. Oocyte and its 1st polar body.

Recombination has occurred, but each cell is still diploid and will need one more division to become haploid. Since most of the lab is interested in the recombination events during meiosis I, this is where most people fix their oocytes onto slides. The percentage of ones that did produce a polar body must be 85%, or else you cannot use any of the oocytes. I have yet to fulfill that, but according the Ailene, the lab tech/'egg queen' it can take people weeks-months to get this perfected to get 85%.

On Friday I practiced collecting some 2 cell embryos from pregnant females (Figure 5).

Figure 5. Mouse 2-cell embryos. Have undergone 1 round of mitotic division, polar body still visible.

The embryos are much more stable than the oocytes, and definitely more forgiving when it comes to time. They are also quicker to collect because instead of looking for unfertilized oocytes still in the yucky brownish slurry mess of an ovary, oocytes have already been ovulated out of the ovary, become fertilized and continue their travel down the oviduct/fallopian tube, where you find them. The oviduct which is a much tougher and whiter piece of connective tissue, making it much easier to see and collect the embryos. This is encouraging since my project involves working with embryos (8-cell).

My animals began getting their BPA dosages and I'll be collecting their embryos on Tuesday and Thursday. I may not be able to FISH them because our lab ran out of the X chromosome probe I would need. I may wait for it to come in, or use another probe to look at something else entirely. I'll have to caucus with Pat and the lab on Tuesday to determine the best plan of action. Only 2 more weeks of my lab rotation!

Come Together

It's the end of the first week of classes and Labor Day Weekend. Well deserved. Classes so far have been mostly review. DNA replication in Molecular Cell Biology and the first 3 chapters of Lehninger in Biochem. We don't have an exam until the last week of September, so it will be ton of information, and our exams for both classes are in the same week, for every exam, which is pretty lame.

Rotation has been the complete opposite as far as requiring my brain power. After a brief pow-wow with Pat on Tuesday, she felt that she already knew what I was capable of, especially after seeing my poster at SSR, and that she was comfortable with me finding my own project. She didn't feel like the end point was necessarily that important, since it's only 5 weeks, but that I get to get a feel for the lab and the people. So of course, I had no idea what to do, given all that freedom. After talking with almost everyone in the lab and freaking out a little, Ailene, one of the research associates, brought up a project that one of the recently graduating students was planning to undertake, but never got around to. Long story short, I pitched it to Pat, and she gave me the green light. In the meantime, I have learned:
1) their kill/collection process
2) how to collect tiny TINY mouse oocytes (eggs) (becoming a master of micropipette making in the process)
3) how to fix embryos on slides
4) #2 and #3 require the odd and ancient skill of mouthpipetting.

Simply put, my rotation project will be exposing pregnant female mice to a short-low-dose regimen of BPA (3 day exposure). I will then collect their embryos which should be in their 8-cell blastocyst stage. I will then use fluorescence in situ hybridization (FISH) to look for aneuploidy where the cells have an abnormal number of chromosomes. The trick to this project is really just getting it done in the 5 weeks I have. I also have to rely on other people to expose the animals since I haven't been trained to that, and the person who does that training just had back surgery--so that won't happen anytime soon. Im excited to work on this, if it works, and there is aneuploidy in the embryos, it will mean that BPA, even at low doses given only for a short period of time, can cause extreme defects if it is during a critical point in pregnancy. It will be a good chance to impress Pat and the rest of the lab. Hopefully I can pull it off.

Last night was a fun get together at a house some second years and a first year live in. I thought it was adorable that it was painted pink. It was a beautiful house, nice front and backyard, granite countertops, and pretty large bedrooms. There was an abundance of food, alcohol, and fun conversation. And Rock Band. Which I am now a fan of. :)

It's also been a whole week with my new little kitten, Mochi. Which no one in Pullman knows how to pronounce, let alone what it is. sigh. Too ethnic I suppose. It's been delightful. She's adorable, extremely entertaining, and very smart as most cats are. She's been handling her time alone surprisingly well. She had a problem with trying to sleep on my face in the middle of the night, and has learned to cozy up just above my head. She also can become overactive between 3-5am, but hasn't done it for awhile, so hopefully she is settling in.


PS-As you can see, the camera made it.