Peddie EXP 2017

Week 6 was my final week! The time went by faster than I ever could have imagined - six weeks seemed a daunting stretch of time at first, but I realized how short it really was as I started my sixth week at JHU. I wrote up and finished my bone marrow monocyte isolation protocol over the weekend and began my final week by going over the protocol with Snow. I got a clearer sense of what we would be doing, why we were doing each step, and what the overall purpose of the experiment; she answered all of my lingering questions and provided helpful tips for me to keep in mind in anticipation of and in preparation for the experiment. For clarification purposes, my new protocol was called "Bone marrow monocyte enrichment using the Miltenyi bone marrow monocyte isolation kit" (Miltenyi is the name of the company that makes the enrichment/isolation kits). We wanted to look at how many monocytes could be obtained from the bone marrow, as well as test out the aforementioned Miltenyi k...

Week 5 overall was not the busiest, but I still learned plenty of useful skills and techniques, important for lab work! I started the week off with some more familiar territory: PCR! It was not the real-time PCR that we did last week, but rather traditional PCR, a process/method I learned about and was able to practice in Biotech my sophomore year. I was definitely more comfortable with this! I did regular PCR with Monica to genotype mice that various lab members had bred; the DNA samples came from tail snips (literally the ends of the mice's tails) that Monica took from the mice. We ran the results under a machine that had various light settings and that projected the results on a computer screen. I even got a small printed and laminated PCR result sheet! It turns out that 9 of the 20 mice we genotyped were IL-4 receptor alpha flox heterozygous mice, and the other 11 were homozygous for the IL4Ra flox gene. the traditional PCR results I then continued analyzing the flow...

My fourth week at my lab got off to a busy start on Monday! I jumped right in with my mini-project given to me just last Friday, which took me the better part of a long (yet anticipatory) day. I'll take this moment to backtrack a bit and provide some information on my experiment. I am comparing the LD column and the autoMACS bead selection to determine which will result in maximized cell enrichment, as well as which will allow for (n oncontaminated) cell culture; I am also going to determine what volume of selection beads to use to filter out  as many unnecessary cells as possible. If I were to simplify the description completely, the LD column is basically a magnetic field. The user adds magnetic "beads" to the sample and these beads adhere to the cells that have a certain marker (e.g. CD45+ cells); the magnetism of the LD column basically holds the magnetic beads (and thus the attached wanted cells) while letting all other unmarked, non-magnetic cells filter through....

Week two, because I got my animal training approved and all my paperwork done, was when things really began to get interesting! Just a quick overview of the past two weeks: I dissected a mouse, helped Snow perform sacrifices and heart harvests, analyzed cardiac tissue using flow cytometry, and learned about projects that'll begin next week. I was also able to visit the Inner Harbor and Washington DC this weekend, a due recess after a busy workweek. I wrote two protocols with Snow, handled some more mice, analyzed a lot of flow cytometry data, and even got my own mini-project (in addition to helping Snow with hers)!  Things got interesting on Monday. Snow was sick, so I was shadowing Paul and watching what he was doing. One of the mice he was immunizing in the animal room suddenly died in the hood (the sterile area that people work with live mice in). Katie and I took it back to the lab to dissect it to see if we could figure out what went wrong. When we cut it open, we could see ...

Week One of my time at Johns Hopkins University got off to an interesting start on Monday, June 19! My Airbnb is within walking distance of JHU's East Baltimore (hospital and medical school) campus, so I began my first full day in Baltimore with a brisk (albeit warm) walk to the Volunteer Services Office to pick up my official volunteer ID. I left early to make sure that I had sufficient time to get my badge and make it to my lab before 9, even with any directional difficulties! I later got to Ross Research Building, where my lab is located, ten minutes before 9 and met my fellow lab members. Katie, an undergraduate student from Muhlenberg College who is also a summer intern like me, was the first person I met. She introduced me to Monica, the lab manager and technician, as well as to Paul, a postdoc at JHU. The fourth-year JHU grad student I would be working with, Snow, came in a bit later, as did HeeSun, another fourth-year JHU grad student. I met my PI, Dr. Daniela Cihakova, sho...