They're Dead Now : (

All our happy cells have died, in what we like to call a mass suicide... or massive apoptosis... depends on if you are listening to me or someone that knows what they are talking about. Anyway, they killed themselves over the weekend, but some of the survivors did grow neurites, so... Yeah!

THEY'RE ALIVE!

I'll separate my personal stuff from my research stuff now:
Research: I have renamed Rachael (my advisor) because she really doesn't like being called Ratmaster because she resents that she has to kill the rats. So I have changed her name to Splinter, the Teenage Mutant Ninja Turtles master who is a rat. But I think this is better, cause she is sagely like Splinter. She likes this one, I think. She likes Kyles name, Captain Bucket Leg. They have yet to really except the idea that I remember everything, so it was shocking to them that I was telling them about things they had already forgotten about. So that was cleared up.

Splinter did another dissection on Tuesday, getting us a new batch of cells for Experiment #2. And on Wednesday they were still alive and doing well in most cases. We have four different samples.

1. PEG (-) Control
2. Fibrin(+) Control (5F,1T)
3. CoPolymer (5F,1T)
4. PorousPolymer

Cells were surviving well in all but the Porous Polymer, which we are really pondering about, because we got some serious pockets in them. I haven't been all that accepting of the Porous Polymer idea, and I let Splinter in on that on Wednesday. I have serious doubts that the polymer will last long enough to be useful in vivo, because as I see it we are just allowing the polymer to hydrolize from the outside-in and outside-out at the same time, which decreases the halflife significantly. I also think, and Splinter agrees, that the process of extracting the Fibrin polymer from the CoPolymer could be detrimental to the cells. We decided Wednesday that we would do an experiment to check out what these pockets we saw in our day 1 sample of Porous Polymer actually consisted of.

What we saw was big pockets that had all our cells in them. We don't know if the cells were in pockets of PEG or nonPEG, and it makes a HUGE difference. And we need to figure out how these pockets are made, because by our thinking they shouldn't happen. My thinking is that when we are collegenating the fibrin out (our way of making it porous), we are taking out the support structure of the PEG gel, making it collapse in on itself. Who knows... we are gunna look at that.

My current quest is to figure out why our methacrylated PEG is different every time we make it. My thinking is that if we do the same thing every time we should get the same result, or atleast very similar results, but we don't. So I have been thinking about possibilities, but its difficult since I don't do this process, its done for me, so I have to read through our lab protocols and look for research on the internet for anything relating to the methacrylating. The last batch that I used was paculier in that it turned our PEG polymerization solution BLUE! Which is concerning, it was actually funny cause we had BLUE PEG and PINK Fibrin, so I told Rachael that I would take the boys and she could have the girls. Here are some pictures of our Day 1 Results: First is the Fibrin Control, Next is the Co-Polymer, and last is our perplexing Porous Polymer.


Personal:
Well we survived another week in the physics library. It was eventful in that people have started banding together again. One interesting part is that one of the new physics chicks has already identified that I "easily offend people." Which didn't bother me too much because I have been pretty ubrupt with her cause she annoys me, but when she said it she knew my name, and I never told her what my name was. I suppose other people don't get the idea behind my calling people Mr. and Ms. instead of using their first name, unless I consider them on a first name basis with me. Oh, and she doesn't like being called a chick. THAT is annoying, I decided that I'd just not talk to or about people that don't like what I call them. With the obvious exception of Splinter.

NEWS FROM THE KRIPT

The week is finally over and I didn't do ANYTHING at work or school except write a Undergrad Research Opportunities (UROP) research proposal. Damn was that a lot of work, a couple of sleepless nights, but I got it done on time and got it into the office just as they were closing. It was pretty close. Here is what it ended up looking like (Neurite Growth Motivation in Polyethylene Glycol Hydrogels). I had somewhere around twenty people read the introduction to make sure that it was plain enough that Joe Blow on the street could understand it... well actually any Joe Blow ITookAnIntroToBiologyCourse should be able to understand it.

To be quite honest, I think my advisor was more stressed out about it than I was. She did make it quite clear that our success in research was linked, if her stuff doesn't work, then my stuff doesn't work, and vice versa. I don't know how me getting paid has anything to do with it, but she really went the distance in helping me get everything done in a short amount of time. My only major contribution was showing her and Captain Bucketleg how to type over PDF's like on the first page of the PDF file. The sad news of the week is that the dissection for the rat embryonic stem cells didn't go well. Apparently they hadn't been in gestation long enough, so we didn't get very many cells, so it was just as well we took all week typing up these proposals. In other Sad news, we only had one of three new methacralated polymers come out well, which doesn't bode well for us. Somehow Dr. Mahoney and MRK (Short for my superiors Maria, Rachael, and Kyle) are gunna have to come up with a way to make that stuff come out with some kind of regularity. Its really concerning that we can't get a good batch done. Maybe its my electric personality causing interferience... I'll have to talk that over with Ratmaster. But it looks like if we can get this thing working Rachael and I will actually be PUBLISHING a paper on it. THAT will look good on my grad school apps.

On the light side of lab news, we got some new peoples in there! One new Grad student joined Dr. Stephanie Bryant's Group, and one new undergrad joined that same group, and they are both gals. The Mahoney and Bryant group share lab space so I'll see a lot of em. I got to bug them when Ratmaster went MIA for an hour Friday. I got to watch 'em do their first polymerization, I think they did somewhere around twenty hydrogels, thats more than I have ever seen done in one sitting.

I talked to Denny today and she will be going to church in the morning, so I am looking forward to that for some good old fashion fun. Hehe, I went to meet my new therapist this week and had to explain why I like hanging out with older people so much. I laughed at her and said that I was going to have a sign made up and laminated that says, "I like them because they MAKE SENSE!" Cause people always ask me.

My Job Is Fun

So this week ended up being... a little productive at work, if not at school. I have all kinds of file now and have been doing all sorts of reading about Bio and Chem. Holy CRAP do they have a lot of acronyms, I thought physics was bad. And all the ones I learned from the papers endedd up being different than the ones that we use in my lab so I have all kinds of Bio terms gumming up the works in my brain for no good reason. I think it is some sort of plot to keep me down.

This picture is the icon I have on my iMac for my research files from the Mahoney group. I thought it was fitting, but we don't really keep mice in jars of green liquid with transmitters implanted in their heads, we only use rats. And speaking of rats, my adviser... I'll call her RatMaster, until I come up with something more permanent... had a terrible time this week because the dissection of the rats for our cells went badly this week. Good ole Captain Bucketleg cheered her up on Friday by calling the rat company and learning that RatMaster has been ordering the wrong rats, for a long time. Now normally this would be bad news, but she took it well, because it means that it was the rat and not her that was screwing up the dissection. We needed this little pickmeup because our cells were dying like crazy! The above is what is supposed to happen to our gels. First we have a lot of fibrin and all PEG, then we mix em (the third picture) and then we take it out and get the last picture. Now if you are confused because the second and last pictures are completely black, well so was I at first, but it just so happens that we pick up the Fibrin with an identifier that reflects back red light, and the PEG does not. So when we see black screen where we knew we had lots of red before, we say HURRAY we got the fibrin out! So what you are seeing is 1) All Fibrin; 2) All PEG; 3) Fibrin + PEG; 4) PEG(w/ holes) - Fibrin.

So on friday I got to watch while RatMaster used the Pascal Imager which is a HUGE microscope that has an laser fed into it from an even larger apparatus which fills about a fourth of the room this thing resides in. It shines different wavelengths of laser light on the sample, one is a pretty blue I don't think i am supposed to look at but do, and it reacts with the indicators and then feeds the image to a computer where we make little movies or slideshows of the output. In this way we got a three dimensional view of what is going on in our gel... which is not what is supposed to be going on in our gel. Here is what we should see, if we were using Pure PEG. The green are living cells and the red dots are dead ones. We should have saw a lot of green with little green arms hanging out. But we didn't we saw a bunch of red, lots and lots and lots of red. It was like a big red cell graveyard I tell ya. But on a more positive note, we did have something in our sample that looked like a rope, that was pretty interesting. I begged Rachael to take a picture of it for me, but she didn't want anyone to see a rope looking thing in her sample so she wouldn't do it, even when I promised to be her bestest friend forever. That takes a lot of pride. So I keep on learning more and more stuff about the lab, and have even learned how to use excel for something useful! Keeping track of the Squirrel Dissention trends

One Week In The Can

SUCCESS! I MADE 4 HYDROGELS! AND I TOOK A DNA COUNT!

I finished my first week of the semester and it looks like it might not go that terrible afterall. Mrs. AdvisorLady is taking it easy on me, that or she doesn't expect all that much out of me. I don't even have to go back to the lab until Thursday because of this holiday weekend. Its upsetting because I really wanted to get back to work. Yesterday we spent the day mashing up our little pucks of cells and then taking the sonicator (I eventually got Rachael to call it a sonificator) and wonking the DNA right out of there.

The lab is great! I am just tickled pink about it. Everyone in The Mahoney Group has an excellent sense of humor and are very stressed out, which leads to much hillarity. Last time I was introduced to Captain Bucketleg - a grad student awaiting Melissa Mahoney's approval on a paper. Then there is The Human Vortex, a grad student bored with conventional means of mixing samples and resorting to shaking everything she can by hand. Then there is Targus! He and I have the same backpack, and he is new to the lab too. He got there a week before me, but like the others... he is a grad student. Next on the Grad Student list is Cast, who is currently on the injured roster, but still she manages to make small talk with me from the Bryant side of the lab and make a crap ton of noise banging things around. Seriously, I didn't know slamming things on a counter was a scientific technique until I spent time in this lab.

I think I'm learning pretty fast how to do all this stuff. Mrs. AdvisorLady was very pleased when we ran the DNA count and it turned out that my standard deviation in volume between all the sample as small. She said she couldn't have done better, but I think that she would have done it MUCH faster. But from the sound of it, they have had problems with undergrads not being so careful so they are okay with old Eeyore here.

Last, I think Rachael is gunna get me out of Gen Physics 3! WAHHOOO!