Peddie EXP Summer 2018

Going into this week, I was a little bit terrified.  My grad student, being away this week, had left me a list of tasks to complete and some protocols.  I really did not want to have to send a text to her saying, "Please help.  I messed up a lot!"  (spoiler alert: I didn't have to) My first job on Monday was to rinse a primary antibody off of western blots and to then put on a secondary.  I was in charge of four westerns which had different lid covers and were all slightly different from each other.  Two westerns followed one protocol and the other two followed another protocol.  When I put the westerns down, I oriented them in a certain way so that I would not accidentally switch the lids.  I then had to rinse off the secondary and expose the westerns to ECL in order for the proteins to become visible in the imager.  After successfully imaging the westerns, I used the stripping buffer in order to prepare them for the next primary, in order t...

By the end of four weeks, I finally feel a little more comfortable in my lab. Even though everyone was incredibly welcoming and inclusive since the first day, it took me a while personally to be able to relax and offer help when I have nothing to do. I have essentially gone through the whole process of how we collect data in lab including everything from dissection to confocal imaging. My mentor, Anna, was not here this week because she went home for the holiday. As a result, in addition to working on my project, I was able to learn a little more about what everyone else in the lab is doing. Even though Anna and I work with the central nervous system, investigating the spinal cord and cerebellum, most of the lab is involved in the enteric nervous system, investigating small intestine and colon tissue. I was able to observe, learn more about and even dissect some gut tissue from our mice (it's not as gross as you think, or at least it wasn't to me). The biggest adjustment I ha...

My fourth week at the Missouri Orthopaedic Institute has been the busiest and most action-packed of them all! Surgery days, end-of-culture days, harvest days, and an immense number of media changes made for a truly unforgettable week. Although I have yet to receive any knee joint tissues, I have been working extensively with my rat tail IVD "Trucker" model that I mentioned in my previous blog. After harvest, I cultured the IVDs under a load that simulated the compression and vibration that the spinal column experiences while driving in an 18-wheeler. The IVDs were cultured in Flexcell plates and vices that connect to a vacuum system, which forces a rubber membrane to compress the IVD against a plastic insert. I developed and programmed a 6-day regimen that followed legal commercial driving limits and allowed for a 1 Hz/.25/.50 MPa pulsating load in order to accurately simulate the vibrations and jostling that occurs while driving. All three of the plates were filled with a...