Silver Enhancement

Well, I went ahead with my silver-gold staining today anyway. I figured that the expensive reagents from this experiment had already been used, so I might as well prepare sections for EM. This necessitated another 12 hr day, but I think it was the right call. Tomorrow I'll re-run the c-fos staining & urotensin staining to make sure that I didn't just mess something up.

In response to an earlier comment, my birdhouse (oops, it's actually a bird-feeder) looks like this (so far):



I want to paint some leaves & such on it, and I'm finally getting to use the paint brushes that my sister sent me many years ago! I like doing creative stuff, but I never seem to get around to it.

That's all for now - it's late & I stood all day today at the bench. I need some sleep :)

Crappity crap crap

Urgh.... well, my c-fos staining did not work, my urotensin staining did not work, and now I'm not sure whether it's worth doing the EM staining for choline transporter. Another fun day in the lab.

I'll try again tomorrow, maybe I just messed something up - but I suspect that the urotensin antibody I'm using is no good, and I probably did not hit my target either.

At least I got to go home a little early, and even picked up some paint in order to finish painting the birdhouse that I started quite awhile ago.

Return from FL & Another Day off

We made it back to Pennsylvania rather uneventfully, and had a long drive back to the house due to highway traffic & some creative use of backroads. But it was a beautiful afternoon, sunny but cool, so it was pretty enjoyable. When we reached the house, we took a walk through the backyard & looked at our produce.

The peaches are just starting to ripen, the plums & pears too, and the tomatoes aren't far behind.

(Okay, I meant to post pictures of the trees & peaches & such, but apparently I don't actually take pictures, I just think about taking them). Perhaps later.....

In other news, my insane experiment that was supposed to happen on Monday (and was the reason that I went to work all day on Sunday to prepare) did not happen. We were out of anesthetic, & I didn't know where to borrow any at 7 am. I actually did this experiment today (Friday), and it took over 14 hours, so I'm glad that I scrapped it on Monday, since I wouldn't have gotten out of work until well after midnight. Science really stinks sometimes.

Thankfully, all of the adverse event stuff that was happening after my recent surgeries has stopped. DR #11 is alive and frisky after a ~ 4 hour procedure on Wednesday. And my two guys today went pretty well - we'll see if I got the activation I'm hoping for on Sunday. Sorry for the dorky stuff - I'm injecting this peptide into the brain that is supposed to cause increased neuronal firing in the brainstem cholinergic neurons, but so far it's only been published for live animals by one group - and they didn't show any c-Fos staining (a protein that is a marker of increased neuronal activity). This is what I'll be doing this weekend, but keep your fingers crossed for me - I've never done this staining before either! Love you all, & I'll post pictures when I get them. In the meantime:



I figure I can post this because it's not data, just an example of how coverslipping errors can produce cool-looking things. This is an area of my tissue where the mounting medium didn't completely cover the section, so there are air bubbles between the coverslip & the tissue - not good for data collection, but I liked it enought to shoot the image anyway. It's also the background on my desktop computer now.

Feeling better

Once again, sorry for the depressing post last time. But I'm pretty sure none of you are strangers to various forms of social anxiety, so I figured it was a good place to post my feelings. The last couple of days have been much better, although I got a migraine today around 2 pm that is making me progressively more nauseated. Bleh.

Since rebuilding one of our garden divider walls, I haven't done much in the yard. Still have to call an electrician; still have finish taking out the 1st floor kitchen ceiling (and floor); still have to get the new heating system squared away; and put up insulation; and put up drywall. This whole house renovation takes much longer than I thought, but a few months ago I decided to be okay with us not really getting much done on this until after I graduate. Since then I've been much happier; but I still wish I'd win the lottery or something, and just be able to hire someone to do all of this (although it's still difficult for me to deal with calling workmen & get them in for estimates).

As far as work is concerned, I'm still running in circles. I am getting closer to being done with Aim 1b of my thesis (only ~250 pictures to go), and will start Aim 2 once I get back from vacation. It would be nice if this would just work right off the bat, but I've been in grad school long enough to know that this will not happen. And in a strange, sarcastic sense, I'm actually looking forward to seeing what weird technical problem comes up this time. Otherwise, I'll continue to try to carry out the dual retrograde tracing (persistent problems with tracer detection, and survival issues), try to get rid of my extranneous bands in my Western blots (ugh, I don't even want to get into it, + communication problems with that PI), resample my dual fluorescence tissue (I didn't save the images from each separate channel for alot of my sampling and only recently realized that this is giving me false positives, but at least I have actually settled on a darn analysis protocol). Still, I had wanted to be done with Aim 1b last year some time, then start work on all this other stuff. I apparently am continuing my trend of not being able to say no to people when they try to make me do things that don't make any sense. But it's hard to contradict someone who has the power to say you don't graduate :)

Still, with any luck, and a tremendous amount of work, I may actually graduate in 2009. We'll see. Also with any luck, we'll have a finished back porch, and upstairs bathroom, & so I will actually be able to throw myself a party (though I may be even more stressed out by this than by my thesis defense!)

Shame

I actually hung out with some of my fellow grad students tonight, and we ended up coming back to my house for a tour of the back yard, since they were dropping me off anyway. I had a good time, but still felt uncomfortable, and the feeling really only grew after they left. I almost always feel that I said something wrong, or just that I am wrong, after spending time with people that I work with or go to school with. And I am always uncomfortable with having them over to the house (with only one or two exceptions). So, after they left I was feeling pretty crummy, and I put my finger on it. I'm ashamed of my house, my neighborhood, my life. This isn't something that comes from any specific person (and my three friends are good people, and I'm sure that they don't think or feel any negative things about me because of my economic position). Rather it's just the awareness that I have that I don't have the same things that they do, that I'm part of a different class, and that no matter how much time I spend in their schools or workplaces, I will never actually be part of the club.

I know that this is not some club that they all decide to keep other people out of. And it's not like I would really change my station in life - I like our house and our neighborhood, and I'm more comfortable with D & our neighbors & friends from the same walk of life than I am with anyone from work/school. But at work, I'm an outsider, and a closeted one at that. With some people, it's fine & really not an issue - I can just keep my personal life personal. But many of the grad students & professors have circles of friends that are entirely internal to the academia, and I just don't fit in there. I felt that way at college too; I guess here it's a little less conspicuous, but it's essentially the same feeling that I had then - when I had to work, and I spent alot of time in Plymouth with people who weren't in on the college thing. I feel like I'm straddling two separate worlds. My involvement in school, etc., kind of inhibits my relationships outside of that arena, and my life outside of school/work inhibits those relationships at school/work. And I feel bad that I relate to people only in a very superficial way - small talk, or discussing pop culture or something. Plus, there are other people, my boss included, who obviously have a problem with my not wanting to discuss my personal life with them, or invite them over, etc. Yet these are precisely the people that I feel would be most uncomfortable with the stark differences between my life & theirs, and I fear that if they knew anything about me, they would realize how much I don't belong around them. Which really makes me miss Boston & Tufts/NEMC - I just don't remember feeling this ashamed of myself when I worked there, and I was much less responsible & grown up there!

So, starting now, I am no longer going out for beers with people after work, because it just leads to this whole shame/awkwardness spiral. I will work late, go to the gym, or walk home & cook dinner for D & me. I will really, really, try hard to quit smoking (at least enough so that I don't do it in front of other people). And I will stop feeling like I have to make lifelong friends out of people with whom I have very little in common.

Sorry for the depressing post, but I just had to get that off my chest.