This past week has been incredibly busy. I spent Monday morning at the Shiley Eye Institute, shadowing Dr. Zhang as he visited his patients, performing eye exams. Using a slit lamp microscope, I was able to examine structures located in the front of the eye, such as the cornea, iris, and lens. Throughout these exams, I could see how devastating the loss of vision really is. Most of the patients that had an exam suffer from diseases that currently have no effective treatments available. From the different eye conditions that I have encountered, Age Related Macular Degeneration fascinates me the most. This disease results in gradual worsening of ones vision but does not lead to complete blindness. As the Macula (the center part of the retina) degenerates, an individual will lose focused central vision. In most cases, there is nothing one can do to restore the lost vision.
For the rest of week, I was busy inventorying the lab’s -80C freezers with the rest of the volunteer crew. Although sampling the inventory is a straightforward task, it requires a lot of time and is a rather slow process. In order to prevent the samples from melting once they leave the freezer, we use dry ice (solid Carbon Dioxide), to keep them frozen. Since dry ice sublimes at -78C, it keeps the samples cool while we enter them into the excel spreadsheets. This in turn keeps sample quality, which will hopefully ensure good results later down the line. I felt somewhat like Alice when she enters Wonderland –or even better Lucie as she finds her way into Narnia – since when I open the freezer doors, I see shelves and boxes covered with snow and ice, which is so incredibly different from when I walk through the hallway where the freezers are stored as the freezers produce a boat load of heat. The inside of the freezers look like winter as the temperature is at such a low level where it will completely freeze all the moisture the air contains. All of these freezers, except for the -150C freezer, which uses liquid nitrogen, run on electricity; it’s incredible. When we finished for the day, we usually liked to put some of the remaining dry ice into water that contains soap, as the dry ice, which immediately begins to turn into gaseous carbon dioxide, will form these amazing co2 bubbles. It’s an odd way of having fun, but that is okay. Depending on how many work on these freezers, it takes somewhere between 2 days up to a week to record the different pieces.
Apart from inventorying freezers, I have also been doing more cell work. A while back I was given some pituitary cells to tend to. Now that I have reached a good cell count, I have passaged them into a twenty-four well plate and have begun treating the cells with puromycin, an antibiotic that is fatal to cells (it causes the disruption of the ribosomes). The purpose of this experiment is to determine the lethal dose (ug/ml), so that when I infect the same cell line with a lentivirus that carries both a puromycin resistance gene along with a gene that will give the fluorescence GFP all the unaffected cells will die. After a couple of days of treatment, only the fluorescent cells should remain. If not, then the dose of puromycin isn’t effective.
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