GAWD! Part 2

Welp, quite simply, everything sucks right now.

In lab, all of our polymers are screwed, so we cant do anything until we make new polymer, and it kinda sucks since we cant get our polymer to last longer than TWO weeks... GRRR...

GAWD!

Ok, so I tried to redo the tests we were running on Friday, but it screwed up AGAIN! I don't know what is up, the stupid stuff is only partially polymerizing! Last time it was polymerizing toward the bottom only so we thought we needed to fix our method of mixing the samples together, ok, so we did that. We even injected it into the syringes differently. We ended getting an evenly unpolymerized gel... GRRRrr. So our new idea is that the polymer that we are using for this test is shot, I used the same one both days, so hopefully it is that because we have a crap ton of that stuff, but only so much of the other ingredients. I'm mad because I did this thing so many damned times and I am sick of it. Its okay if it works, but it is crushing when it doesn't.

On top of that, I think I killed our Experiment 3 SUPER DOOPER COOL CELLs by putting in cold sollution, we will see if they died tomorrow, but as things have been going terrible, I do not hold much hope. It was sooo stupid, I realized what I had done the second I took the little bastards out of the hood and looked at my empty vial, '@#$% I didn't warm up the solution... AHHH! Get the little bastards in the incubater!' Then I had to go tell Rachael what I did, I felt ashamed.

And on top of THAT! I dropped my IPOD and broke it!!! GAWD! But its okay now, I went and bought a new one, thank god I got the grant!

I am so smart-- SMRT... I mean SMART

Lots of things happening this week. Splinter was perfecting her photography skills, I was perfecting my standing around looking pretty skills, and I got a GRANT!

So we will start off with the grant. I was able to get the Howard Hughes Medical Institute's Bioscience Grant for laboratory-based biomedical research. This is really great because if I am planning to do Tissue Engineering for grad school and chuck the physics (well, just not major in physics), so it will be great to put on an application for a school that does a lot of medical research... like say... hmmm.... Johns Hopkins! I am really psyched about this grant, cause Rachael and I are used to being turned down on achievements like this so its fun to finally feel that accomplishment. The only downer is that other people are used to it so they just look at you funny when you are celebrating. So with this grant comes some extra responsiblity, but because I am taking an EBIO writing class I'll probably have my entire paper for this project done by the end of this semster, much earlier than when it is actually due. And since we are pretty much prepping for publication already, I don't think there will be much of a problem coming up with really cool things to write about.

Next! So about this publication thing. Right now we are in the process of verifying all of our data, which means Rachael is taking a lot of pictures of our cells on the big expensive confocal microscope, while I am off preparing samples for photography and other testing. Last week we prepared for immuno testing, which was when I ended up fixing up the Anseth deli slicer. Those will be ready for imaging monday, and hopefully we will get some good pictures out of that, it took enough time to get the stuff ready!

Friday, was very disappointing because the samples we were going to use for florecent tests didn't come out right, and we don't know why. We have some ideas, I personally think that I screwed the polymer up last week when I forgot to put it back in the fridge after I had used it, Rachael remains adimant that such a situation would not effect the polymerization. The next idea is that our fibrinogen is bad, which would suck! Lastly, our mixing method was off... who knows, it took for freaking ever so and I was all pissy when we were doing it, I don't look forward to doing it again.

So we are going full steam ahead with publication, and rightly so, because if we got what we THINK we got, a crap ton of neurites growing in microchannels we made! We are really excited because if this is true, we made a BIG leap in our research and thus making treatment of NDD's a nearer possibilty. YAY!

Andy

DAMNED SHREDDER!

Welp, good things are happening in the lab if not in life, but as many pictures that we have of our samples... I cant find a darned one of em that is cool without a 3D projection! So I'll probably take some pictures on friday throught he normal light microscope and post those. Hopefully the cells last that long. In other lab news.. Splinter and I are looking at publishing our findings so we are currently preparing for our next set of experiments. It is going to be the same experiment, only we will be doing it much more exactly and with lots more documentation. Today Splinter and I practicing for the pictures we will be taking to include in our article. To do this we had to use a CryoSomethingorother and take micrometer slices of our hydrogels so that we can MANUALLY do what we have been doing with the confocal... looking at our gels layer by layer. Also we are gunna be doing immuno tests on it.

My big adventure today was to try do these gel slices without Splinters aide, it was a disaster because the machine broke 5 minutes after she left and apparently no one knows how to use the stupid thing. So after an hour of dinking around somone finally let me in on how the thing worked and then I was able to maximize the output of the slicer getting really good slices of gel. I totally left that damned machine running at 200% from when I found it. When Splinter got back I had to show how to do some of the things I had found out so she could use em. Its really fun to try and impress Splinter, because she is always really enthusiastic about our project and if we figure something out that helps it along she gets really excited. Excitement is something you don't see in the sciences too much.

Andy

We got new Babies and They are growing Neurites!

These are some new pictures we took of the Day 1 samples on September 27. They look really good and we are very optimistic about them living through the weekend unlike the last suicidal bunch.

I'll Explain what you are seeing here. First off we have the PEG Control. This is the hydrogel consisting of pure PEG and rat embryonic stem cells. I think it looks a lot like cement.

This picture is of our Fibrin control. This is a sample that consists of 1 part Thrombin for every 5 parts Fibrin, which means its heavy on the fibrin so we get an idea what the cells would look like if they were growing in a fibrin enriched environment. This is important because neurites are seen extensively in fibrin samples, so if we can beat this we will be very excited!


This is one of the co-polymers with PEG and Fibrin. I think this is the 25 parts Thrombin to 250 parts Fibrin. The reason you can't see crap in this sample is that there is a lot of Fibrin still in the sample, after a while it will get lighter as the cells consume the fibrin. Until then we will be taking pictures of this nonsense. The cells in here look really healthy though.


Ok, here is the interesting stuff. Splinter and I had a talk a while ago about the practicality of using big pores to small pores, so we decided to make two porous samples, one with HUGE pores and one with the minimum pore radius we were willing to use. Splinter says this one is mine: Limited Impact Porous Gel 1T to 5F. She is really excited about this sample, even though I think hers looks better than this. The good news is that we think we saw neurite extension on Day 3, Friday September 29th. Which is HUGE cause we don't even
have much in the fibrin samples.

Ok, here we see Splinters sample Max Porous 25 T to 250 F. Here you can kind of make out a grain structure in the gel, which is really cool, it looks like the pattern you would get in a chunk of wood. It is really exciting because we are doing all this to make channels for neurites to extend through and here we can definitely see the channel structures. I think this is gunna be our winner if it doesn't degrade to fast. Hopefully it will be around on Monday!