Handkäse mit Musik
Pictured here is La Pequeña Flora, The Little Flower. It’s a church tucked away in a residential area that I happened upon. There are tons of old religious structures in San Antonio, I thought I would share my favorite one with you guys. Touring this was better than seeing the Alamo in my opinion. This week was quite eventful, with a human brain dissection and the chemicals for the ELISA kit I’m working on coming in. I also want to give a huge thank you to all of you sending me letters, the mailman here now knows me by my name due to the abnormal volume of mail I receive that isn’t just bills. As I finish off the final steps in the newest protocol (fingers-crossed) I wish the rest of you much luck with the projects and endeavors you’re tackling this summer. Let’s go Deutschland.
The human brain dissection happened this past Wednesday, and for the first time in my life being short didn’t get in the way of me being able to see what was going on, being explained, and demonstrated. There also wasn’t a crowd, so that coupled with the fact that the person doing the cutting was about my height was the reason I had a great view. I’m not sure how every brain examination is carried out, but this one started out by inspecting the dorsal and ventral sides of the brain. This was followed by taking a look at the rostral and caudal ends. Satisfied that the brain was up to par – no missing chunks or signs of rough extraction from medical examiner(s), aside from olfactory bulbs and other delicate parts which tend not to make it through the process – a single sagittal cut was made, exposing a very obvious cingulate gyus, corpus collasum, fourth ventricle, and fornix. The rest was a fleshy mauve color containing the other less distinct structures. Most brains that are displayed to the public have been, or still are, wading in formalin. This brain had no such treatment and was very fresh. Fixing tissue using this process damages and/or modifies RNA in tissue samples, which as you can imagine interferes with measuring any effects from administered or abused substances prior to passing. To avoid this, the brain was kept as cold as possible and was much squishier. The sagittal cut was approved by the pathologist and coronal cuts were made moving from rostral to caudal, i.e. front to back. After this lengthy process and examination, the left and right sections were labeled and flash frozen (methylbutanol and dry ice, if you were wondering…) to keep until a later date.
Figure 1. Dissection table. This tabletop can be set to - 4°C, so I’m told. This keep the brain stiff since it isn’t fixed, but more importantly continues to slow the RNA decay in brain tissue, which is what will be measured in certain targeted regions of the brain related to the psychopathy study being conducted.
I’m happy to tell you that after patiently waiting, I am finally seeing the results I actually want to and think I should be seeing. Emphasis on the latter. This has happened only once though, so don’t get your/my hopes up just yet; these things only count if they are replicable. After much discussion with my supervisors (some deity bless these people, I don’t know where I’d be without them), I decided to play around with the Forskolin concentration. This idea has been modified to also vary whether or not samples are spiked with ATP (+ATP/-ATP). The wells that are not given ATP will serve as controls since cAMP won’t be produced without ATP present (I say with bated breath). I’ll keep this short so as not to excite you and myself too much. An indefinite hoping.
Figure 2. This is what success looks like. Pay attention to wells B4, B5, C4, and C5. These four wells contained lymphocyte samples, Forskolin, and ATP (+ATP), so they were able to produce cAMP which competed with the dye reagent kicking it off the membrane at the bottom of the well, hence the lighter color. In wells A4 and A5, no ATP was administered (-ATP) and the dye was able to remain locked on to the membrane receptors at the bottom. The entire explanation of this well-plate would fill an afternoon over coffee and biscuits, so I’ll leave at that for now.
This just sounded really good.
What I have learned or been reminded of so far:
1. Dolphin and human brains are astonishingly similar. I spent an afternoon looking at different brains.
2. I am reminded that formaldehyde is a carcinogen. I always (choose to) forget that.
3. Hemingway would have made a bad scientist. I swear he wrote the last protocol I’m trying to follow. Here’s an excerpt of an excerpt:
“Pull out filter sheet before degas.
May need to be tapped twice, but no more.
Doesn’t stick – wet.”
That’s all I have to work with at times, sometimes even less. Cherish your Intro Chem101/102 lab manuals and Bio binders, they’re worth the $5.