WISE Journal Entry #9

Name: Leah Gerald

Entry: #9 (3/13/2012)

WISE Week: 5

Written on Date:3/15/2012

Mentor: Sagar


Today was a somewhat slow day. It was slow because there was a lot of very specific things that had to be done, so I wasn't able to actually do anything with my own hands, so I spent most of my time watching. Nathaniel and Sagar were both working on separate parts of the same experiment. Nathaniel was prepping some proteins to act as a sort of "control" for what Sagar was doing, and Sagar was working on extracting proteins from 2 different cell plates, one with YAP (the gene that controls a cells ability to stop or start making more cells) and one with YAP knocked out. Each cell plate had an array of different dilutions of a chemical to see how it will be affected. Later, once the cells were all properly treated, he would test the protein levels in these cells in comparison to what Nathaniel was doing. Nathaniel was basically setting up a standard curve created by the DNA of the same cells, but without the chemicals in it to compare. To see the difference, since we cannot see DNA, what they do is compare the proteins the DNA creates. YAP makes a specific protein, and they can tell if they were able to actually knock down YAP and to what extent by looking at the proteins created. to see the proteins, they use a process called PCR (polymerase chain reaction) that replicates the DNA enough times to create enough proteins and a visible difference between the different amounts. I didn't get to see all of this, because I left before they were able to see the proteins, but I did get to watch most of the progress. It was fairly slow because it was a lot of pip-petting and very specific, small measurements that needed to be made. near the end, they also had a mouse they had infected with the cordoma cells (cancerous cells that infect the area of the brain nearer to the spine) that had recently died, and they needed to open it up and extract the cells. Sagar showed me the mouse, and how the cordoma caused the mouses back to become crooked and a large growth to appear on his back, but by this time, it was already 4:30, and I had to leave before the action.

WISE journal entry #8

Name: Leah Gerald

Entry: #8 (3/7/2012)

WISE Week: 4

Written on Date:3/11/2012

Mentor: Sagar


today was a day for watching. According to my mentor today we were extracting RNA. first, we had to take the cells, wash them, and then put trizol on them. the trizol would seperate all the parts of the cell. I got to see it in action under the microscope and it was pretty cool. I could see all of the parts of the cell breaking into tiny little pieces. once all that was done, we had to put a little chloroform in each of the viles, and then put it in the centrifuge. after that, we put a little isopropyl in them, and they were to sit in the freezer for a while, so I didnt get to see the final results. its sad when I dont get to see things to the end, and when I miss things. sometimes I wish i could just come in for a second so I could see the results, because I really wanted to see the RNA.all I was able to actually do that day was wash out the containers, because they had to make sure everything was done very precisely. It was a little reminiscent of my first days, because I spent most of the time watching, and following, and not understanding the humor.

WISE journal entry #7

Name: Leah Gerald

Entry: #7 (3/6/2012)

WISE Week: 4

Written on Date:3/11/2012

Mentor: Sagar


today was a very short day, partially because we left early, partially because I ended up not doing very much. I was in the lab for an hour less, plus my mentor had a meeting, so I ended up listening a lot. the trays I pipetted last week showed an inconsistency that proved to be problematic. we did 2 sets of each dilution, and basically one of the set of dilutions showed two different results, which doesn't make sense, so they had to re-pipette the whole thing. the week before, my mentor asked me to look up trypan blue, and today, I was quizzed on it. trypan blue is basically a dye they use that colors dead cells blue, because the dead cells allow it to pass through their membrane whereas the live cells do not. they weren't using it today, but in the future they will.

WISE journal entry #6

Name: Leah Gerald

Entry: #6 (2/29/2012)

WISE Week: 3

Written on Date:3/4/2012

Mentor: Sagar


today I did not expect what happened to happen. I walked in, and my mentor was dissecting a little mouse on what looked like an operation table to me. I'm a bit squeamish, and I didn't see this coming, so that was a real shocker for me. these mice were part of there experiment though, he wasn't just doing this for fun. they had mice which they inserted some cancer cells in earlier, and they needed to take the brains out so they could look at the results. they hadn't planned on taking them out on that day, but because some of the mice were dying before they expected, they wanted to extract the brains from the still alive mice before they became useless. so that day consisted of a lot of mice deaths, which took a great deal of time. first, they had to weigh the mouse so they could know how much anesthesia to give them. then, once they gave the mouse the anesthesia, Sagar would put them on a foil table in this section off to the side and pin them down. once pinned down, he has to pump saline solution into the right atrium (sorry if I'm wrong) and drain the blood from the left atrium, so there will not be any blood in the brain when they take it out, and they can clearly see what they are looking for. once that is done, they add some formalin so that the cells will preserve and can sit for a while without decomposing and stuff. they had to repeat this process with several mice, so this day was very much a watching day. I didn't want to have any part with the process with these mice, so I didn't look at the mice as much as I could, but every once in a while I'd try to sneak a peak to help desensitize myself. after a while I could look at the mouse without feeling sick, so I'd say that was my progress for the day.

Wise journal entry #5

Name: Leah Gerald

Entry: #5 (2/28/2012)

WISE Week: 3

Written on Date:3/4/2012

Mentor: Sagar

Today was a special day. Like last week, we went see another speaker speak. This time it was about from what I understood how cells tend to try to organize themselves. He used some words I remembered from Chemistry like "entropy" and I was excited I could actually understand that part, but for the most part, it was fairly boring. Many people in the room fell asleep. In fact, one of my mentors fell asleep. It wasn’t because the material itself was boring, but because it was a fairly complex concept that didn’t interest many people there, therefore they couldn’t relate to it. The exciting part about that day was, before we went to see the speaker, I got to work under the hood for the first time! There was some pipetting that needed to be done. They just needed to change the medium for some of the breast cancer cells. Though it was something so menial, it was hard for me. I believe I took almost twice as long as my mentors. Even though it was my first time, I was shocked at how difficult it was. I had been watching them do it every day for a long enough time now; I just assumed it would come easily to me.