Kelsie, Entry #1, Counting Dendrites and Western Blots

So far the first couple week has gone well.  My first day I met my grad student Julie and an undergrad from Wheaton, Kyle, who is also working with Julie this summer and they are extremely nice.  Both were very welcoming and showed me around the lab then I got to start working.  I counted the number of dendrites branching from the soma of many neurons and after I was done with that I got to actually work a little in the lab by making a solution we use for western blotting.  I also got to see the microscope used to look at the neurons, which we use to take pictures, which is what I was analyzing earlier so that was pretty cool to see the machine that took the pictures I was looking at.

Then the second day I got to help run some experiments which was pretty exciting.  We used gel electrophoresis which I got excited about because we did it in biotech and I felt like I sort of knew what I was doing.  I was doing a lot for that in preparing the solutions and pipetting the ladder into the gel.  After the gel was done I helped with transferring the proteins to the membrane used for the western blotting and once that was finished, used an antibody that is the first step in marking the E2F1 to be visible.  That had to sit overnight.  The three of us ate lunch together which was a lot of fun actually because I was able to just talk with them about who they are.  Turns out they both went to Wheaton and when Julie was a senior, Kyle was a freshman and they knew each other because Julie’s husband was the pastor at Kyle’s parish.  I also thought it was cool that Kyle is from California and Julie is from Western PA out by where my sister lives now.  But after lunch we went on a little adventure to pick up rat cortical neurons from another lab which was interesting to go and see more of campus as well as another lab building.  

Letting the membrane sit overnight, I came in on the third day and first went to a presentation on HIV and the efficacy of treatments and that was really interesting especially since, at Peddie, we had the presentation on combination therapy for HIV.  I got my PennCard so now I can enter the building by myself in the mornings and be able to go right up to the lab.  Finally, returning to the lab I removed the antibody from the membrane and did three washes before placing the secondary antibody on.  During the hour that had to sit, I finally was able to meet my PI, Dr. Kelly Jordan-Sciutto, because she had been away for my first few days.  Unfortunately she had to run to a meeting but at least I was able to introduce myself.  Finally we all ate lunch and I was able to talk to Kyle some more and he was asking about my soccer and the Women’s World Cup.  We ran through the second set of washes and once that was finally done we could use a substrate that would help make the E2F1 visible once the film was developed.  The three of us went down to the dark room which was quite discombobulating at first since there were only two red lights and my eyes had to adjust.  But it’s just like the developing red rooms used to develop the old film, like in the movies.  We got some good results on the film so that was exciting for me since it was the first Western Blot I’ve done.  

Thursday when I went in I got directions from Julie since she had to catch a flight to go to a wedding.  Kyle and I did another Western Blot with another set of antibodies for the E2F1.  During the second wash, Dr. Jordan-Sciutto called me in to talk to her about what will be happening in July since Julie and Kyle will be leaving.  She gave me some grant proposals to read during the waiting periods.  I finished the last wash of the second set and ate lunch and again just talked about soccer and he plays baseball so we talked a little about that too.   Finally, all the washes were done and we could go to the dark room to see any results.  Unfortunately, the machine was turned off or we turned it off so we had to wait for it to warm back up again which took about 20 minutes where all we could do was talk.  Seems like we’ve all talked a lot J.  But we had to develop a few different films attempting to get a good, clear image.  That was the last I did.  All in all, it was a good first week and it got better as it progressed so I’m hoping it keeps going up from here.