Tuesday's Day in the Lab

Tuesday I went to the lab for my second s&h. This one was easier, both emotionally and practically. I did a good job, all my practice brains came out beautifully. U asked me to take out an experimental brain for her (she needed to check the placement of a cannula), which I thought I screwed up but the gouge in it was so large that I've determined it must've been from something besides me. I'm good enough now that I would've noticed if I'd made such a huge mistake.

This week:
Animal Care Training
EH&S Training
More CryoStat (two more brains?)

Today's DIL Report

Today I went to the lab to use the CryoStat (it turns out it was just locked last week, and it just needed the "lock" button pushed). I sliced an entire brain. It was one of the four from last week (two were mine, two were U's) and it was definitely one of mine -- the olfactory bulb was quite messy and folded under the cortex. There was also a large slice through part of the cortex which C thought was from uneven freezing.

Also! C came with me, which was awesome. She graded papers at the desk next to the CryoStat while I sliced and looked over my slides. She says that they're very good, even amazing considering it was my first time slicing. Every set of slides she'd look over my slices and show me landmarks from the mouse atlas. That was the best part of the day, looking over my slides with C and her teaching me what on the brain corresponds to what in the atlas and some of the functions of certain major structures.

As for the actual slicing, I think it was pretty good. The olfactory bulb messed up the first few sets of slides and I had some serious cracking issues, but they couldn't be resolved by adjusting the temperature which means it was most likely a problem with the brain itself. I got a lot of a sort-of "venetian blind" effect, where the slices had a series of parallel horizontal cracks. Both C and U said that it's not desirable but also not abnormal and that there's really nothing I could do about it. Looking at the slices, it didn't seem to mess them up too badly.

At some point I'm going to bring my camera into the lab and take pictures of my slices so that I can look them over enlarged, digitally, and compare them to my later slices (as well as include them in my DivIII portfolio).

Next Tuesday U, A and I are going to harvest more brains, which is great for getting me more exposure to animal sacrifice. I definitely prefer slicing brains to harvesting them. That said, I still want to get it perfect. I want my brains to look like this every time:





In other news, I got a note from Laurie today requesting a letter for Rayane's file (and I assume one will come for C's too) so I better get on that. I also need to put my DivIII on TheHub so that S can read it. So, list!

To-Do by Friday:
TheHub DivIII Proposal
Rayane's Letter
C's Letter

(Backdated Entry)

On this day (since I am writing from THE FUTURE) I went to the lab to learn to use the CryoStat from Se and to harvest brains with U. There were no practice brains, so Se couldn't do more than walk me through the process and mime the actions. When U showed up we harvested brains for practice from some male voles.

I had a severe reaction to watching the animals be sacrificed. H has suggested I start a journal about my feelings towards this subject so I can remember everything. I will be starting that journal but it will be password-protected.

Besides my emotional state in the lab, the day went rather well. I extracted four brains, of which two were not so mangled that I could practice slicing on them. My first brain took about 15 minutes, my final brain was less than 5 min. U says that my learning is very fast and she thinks I am talented at removing the brains.

I need the most work with the olfactory bulb. The skull is very thick over it so it's difficult to remove without harming it. It's also gooey-er than the rest of the brain, so it gets squished or disoriented much more easily. I know what I need to work on.

Moving along very nicely

I've now been to two lab meetings and had two private meetings with U. It looks like I'll be picking up where her former student left off, doing the in situ hybridization of vole brain slices. I read the protocols over the weekend and it looks like 50/50 stuff-I-understand and stuff-I-don't. Hopefully that's a good ratio.

This week I'm going to meet with Se, one of her graduate students, who will teach me how to make slices in the CryoStat. U said she'd meet with me before that to harvest some male brains for me to slice.

The project is moving along nicely. My only problem right now is that I'm squeemish about killing the animals.

Met with U

I met with U at her lab meeting on Tuesday. I emailed her to find out when she is available to meet with me personally, but she has yet to respond. It looks like a good situation, though, and I'm excited to work in her lab.

I don't think I'm going to know what I'm doing fast enough for the CBD proposal, unless I write it more generally, but then I will probably not get any funding.

Actually Updating Helps

Thursday: As I mentioned a post or so ago, I contacted companies and analysis of samples is very expensive ($700-1,000 depending on the company). Rayane contacted Axys Environmental and spoke to one of their analytical chemists, who explained why it's so expensive. It is standard to spike samples with radioactive PBDEs, as somehow this helps the chemists figure out what the baseline background levels are (or maybe a ratio or something). Radiolabeled PBDEs are expensive, about $1000/mL. This is necessary because the risk of contamination is so high.

But how high is it? The chemist told Rayane something like, "A new couch in the building will contaminate your samples." So, basically, the work is so sensitive that it wouldn't be possible to get good analysis on campus, so I'd have to send my samples to an external lab, which is just prohibitively expensive.


Friday: C asks me 100 questions about my feelings on animal research. Apparently there's an opening in Professor U's lab at UMass. Prof. U works with prairie voles, which is something I looked into for my DivIII when I was in C's Comparative Animal Physiology class. Prof. U had a senior doing a project that is somehow integral to her research, but he can't do it anymore so she needs a student to pick it up.

I don't know what the project is, but I even remember talking to C about Prof. U two years ago when I was interested in prairie voles and vasopressin/oxytocin. I am very excited to find out what's open for me over there.

The bad news is that Rayane will not be on my committee if I'm working with animals, so I have to rebuild it. C will be my chair, and she's already shuffled some of her DivIIIs around so that she can fulfill that. I think I'll ask S if she wants to be my member, once I find out exactly what I'm doing.


Assignments for break:
Email Prof. U
Read Prof. U's work and any other papers she recommends
Write for C and Rayane's reappointment files
Breathe

The Return Call

The rep from Eurofin called back. I missed her call, but she left a message which was very slow with lots of "ums" and "uhs," but she said the PBDE analysis was only done on fish oils and that it cost $750 per sample. Then she said her number at the speed of light and hung up.

Is this the end of our hero? Stay tuned 'til next time...

Start of Funding Proposal (and: harrassing corporations still gets you nowhere)

I have a draft, though it is poor and both too detailed but not detailed enough and very reflective of my general cluelessness of how to write a funding proposal. There are some good ideas, though, so it is something to work with. It can be found here.

I spent a good amount of time (about 2.5 hours, for anyone keeping track) on Thursday looking for chemicals. I seriously harrassed Chemtura (Great Lakes is now a part of Chemtura) to find PBDEs, but nobody high on the corporate foodchain was available and everything else directed me to customer service, who said they didn't have PBDEs available anymore. I sent 3 emails, no response yet, but that's expected. Maybe I'll get something back, who knows.

I also found this site, which I have to go through company-by-company.


EDIT: This is noteworthy. re: Sample analysis.

I have contacted Lars Reinmann about PBDE analysis, and I also called their customer support line, for which I should be getting a return call any second now....