Peddie EXP Summer 2018

As I was wrapping up my time at the lab, Claire was also wrapping up some of her work to present to her thesis committee.  My seventh week was a continuation of probing western blots with different antibodies after extracting protein from the heart, lung, and brain of each of the mice which had been genotyped before.  This allowed for the comparison of heterozygous, homozygous, and wild type mice.  The results from the antibodies were unexpected, leading to some confusion.  Thus, we had to go back and redo the genotyping in order to confirm the original conclusions from the gel electrophoresis.  The genotyping was correct, but there were a variety of other factors which could have led to the unexpected results. My 8th and final week was spent doing some lab work, but mostly reading some primary literature in order to prepare for my poster.  I took some notes and discussed what I read with Claire in order to gain confidence in my knowledge of the topic. ...

I began week 5 by using the microscope to view the cell plates that I had previously stained.  It was really exciting both to have results and to also see that I had followed the procedure correctly.  I was a little bit nervous that something that gone wrong along the way and that the microscope but be unable to see the neurons, but I was able to view the red, blue, and green stains.  I then had to set points in the center of each well and then focus the image and change the brightness, in order for the microscope to, hopefully, capture.  However, I was a little unlucky and had to keep restarting the process because the microscope kept crashing, and was not able to take the 25 images needed of each well.  Instead,  Claire was able to take a single image of each well by zooming out, leading to a larger picture.  We were happy to find that the results were as expected, with cell death occurring in wells in which it was predicted, and the cells in the veh...

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...

The last two weeks have been slightly hectic.  My first week in the lab I became used to the easy 7 minute walk from my apartment to the Schattner building.  However, due to the need to go through some mandatory training for Penn, I was required to take a 17 minute walk past Penn and past Drexel to reach the Environmental Health and Radiation Safety (EHRS) office.  This training session covered basic, common sense rules, such as "Do not eat in the lab" and more specific guidelines for chemical safety and biohazards.  I was glad to find that I already knew much of what was explained during the training, thanks to having worked in a fume hood before for chemistry and having had some hands on experiments in my previous science classes. On Tuesday of my second week, Dr. Peretz, Ms. Cozine, and Ms. Terhaar came to take all of the EXPers in Philadelphia (except for Wendy) out to lunch at the White Dog Cafe.  It was really exciting to see some alumni and catch up with...