Just Keep Swimming!

This week has been absolutely crazy for me. I had three exams and final project deadlines are quickly approaching! This week in lab I have worked on trying to come up with an idea for obtaining brain slices that are thin enough for me to see proteins but strong enough to survive the staining process. I started by selecting three brains that are positive for frontal lobe hematoma and three brains that do not have any visible hematoma. I am not sure that there is any significance to this but, I am curious as to why approximately 40 percent of the brains in an unopened bucket are positive and the rest are not. I then used a coring instrument to obtain three samples from each brain and placed them in the -80 degree freezer for 24 hours. The coring process was not much fun, the preservative smell and the close proximity needed for an accurate sample left me feeling a bit nauseous. Perhaps I will do only three at a time for my next coring session. Next I very slowly  made slices using my makeshift microtome. I then took these slices and stained them using crystal violet. I have been able to slice them extremely thin but once I start staining them they disintegrate, littering the sink with brain matter which is a bit gross.  The problem with such thin slices is that they thaw once they hit the room temperature microscope slides. Perhaps freezing them again before staining along with the microscope slides will help? I also got to spend time discussing projects with some of my S-STEM colleagues, celebrated Matt Hills birthday, and tried to offer a warm welcome to our newest STEMmers who were busy with their unknown microbe projects. All in all, it was a great week and I am excited to see everyone's presentations in the very near future!

Hematoma Brain

Normal Brain

S-STEM Field trip

Last Friday I had the good fortune of my first S-STEM field trip. We went to Dreamy Draw park and I got to hike and experience geocaching for the very first time with some pretty amazing people. Thank you to our fearless (and extremely knowledgeable) tour guides Matt and Josh. I learned about femoral drainage and the mining of mercury that used to take place there. It was a much needed break from this stressful semester, and I am so happy that not only has S-STEM been an amazing experience, but I did not realize the bonds that would be formed from spending so much time together in the lab.

This is a pic of Matt and I attempting microtome #2

The search for a new brain

This week has been an exciting eek for me in the lab. I have successfully created a microtome! Ok, it is a very archaic design, but functional nonetheless. I used a 20 cc syringe, a brass corer (size 7), and a blade from a box cutter. The tip of the syringe was removed using a box cutter. After much measurement I realized that the diameter thickness that I wish to acieve just happens to be the distance between the cc markings on the syringe. Matt Hill used a saw to create an opening for my blade. the brain is cored and placed into the syringe. a smaller corer is used to plunge the brain down the syringe until it is pressed against the blade resting in its position within the cut in the syringe. when the blade is released the brain is moved ever so slowly forward and the blade slices through it. This little device has created some adequate samples for staining (using crystal violet) so I am very happy with it. However,  the lego microtome is still in the process of being built (thanks to Matt Hill) but is not quite finished yet.  The first brain that I sectioned was chosen due to the presence of a hematoma on the frontal lobe and a decreased frontal lobe size. This brain contained proteins when stained and viewed under the microscope. However, frozen brain tissue is much easier to cut so I placed it in the -80 degree freezer so that it would be super frozen. Well, when I applied the pressure to it in order to core it...it exploded into pieces which flew all over the bench and floor. although impressive it was really unpleasant to clean up (it defrosts quickly in such small pieces.) Now brainless, I picked another brain randomly from the bucket. after getting a sample under the microscope I quickly realized that it did not contain the proteins that I was looking for. I chose another brain randomly and again obtained negative results. I began searching the buckets for a third brain vowing to be picky this time to find one that shared the same characteristics as the first brain. I found the perfect specimen at the bottom of the bucket. Unfortunately I ended up dipping my leather banded watch in the process of retrieving it, which my fellow STEMmer Jeremy found extremely entertaining. This brain is positive for proteins. I am not sure if this is coincidence or not but it is defiantly interesting.

Beta amyloid present in brain tissue.

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0CAYQjB0&url=http%3A%2F%2Fwww.scbt.com%2Fdatasheet-53822-beta-amyloid-20-1-antibody.html&ei=Kk9cVNP1MY72iQLCn4G4AQ&bvm=bv.79189006,d.cGE&psig=AFQjCNGoJW91fjtiLzgLVGozUWqAPsl6bA&ust=1415422087904595