Peddie EXP Summer 2018

Time really flew by! I'm wrapping up my work here in the lab. I'll miss everything about this summer so much--even washing and rinsing my membrane six times before imaging! These past few weeks have been very productive. I've worked on a lot of Western blots, gave more presentations at our lab meetings, and presented my final poster alongside Blake. Its been super busy and exciting, its sad that it has to come to an end. I'll include some random bits and pieces from recently: Preston took me to image some organoids on the microscope. The pictures are really cool. Its interesting to see the diversity of organoids from cells of the same sort. Basal and luminal-derived organoids have different morphologies as well, which is pretty awesome. Its amazing how these three dimensional tissue models can retain their identities in culture like this. Luminal-derived organoid I also learned how to quantify my Western blot data. The images obtained from performing Western b...

At the start of my fourth week, I was given an independent project to do along with an undergraduate student there, Sarah. Our job would be to perform immunohistochemistry (IHC) on mice brains to delineate regions in the striatum, a region in the brain. IHC utilizes the specificity of antibodies to attach to epitopes on specific antigens. This allows certain cells of a tissue section (with a certain antigen) to be “selected” by a certain antibody. By adding a secondary antibody with a fluorescent tag, target cells can be selectively labeled, imaged, and analyzed. We sought to mark regions in the striatum based on cellular count intensity.   The procedure begins with perfusing the mice.   This is a gory procedure that involves anaesthetizing a live mouse, pumping out the blood from its circulatory system, injecting a preservative solution into it heart, pumping this solution throughout its circulatory system, chopping its head off, and dissecting out the whole brain. This...

These past two weeks have been amazing (and hectic)! My Time in the Lab: I am beyond grateful for everyone in my lab—they are all so helpful and it is truly a blessing to have so many great mentors! This first month in the lab has been an eye-opening experience for me on so many different levels. Immersing myself into the “real world” of science through research has not only confirmed my passion for cellular biology but has also furthered it. I am so fortunate to have the opportunity to participate in exciting and ground-breaking cancer and metabolism research! In my third week, I gave my first lab presentation in front of Dr. Goldstein, Preston, Johnny, Blake, and Ana. I was a little nervous because I didn’t want to present any of Preston’s and my results incorrectly, but I felt that discussing our work with everyone made me more confident in my role in the lab and honed my presentation skills. Dr. Goldstein emphasizes the importance of being a strong presenter in researc...

Week 2 and 3 My second week largely involved further familiarizing myself with the lab, its members, and the cellular analytic work I was tasked with. I continued my work with the image processing of brain slices to highlight the contrast between cell bodies and the “background noise” (while maintaining the original pixel data of image). This was in hopes that we could use some sort of automated way to count all the cells. Although quantifying the relative intensity of the pixels of cells was a somewhat viable method to “count” the cells, the ultimate goal was to actually count the cells. I took a stab at counting the cells in one brain slice – 1181 red cells and 381 green cells – definitely not fun and this only motivated me further to find an alternative method. After some research, I then turned my attention towards a machine-learning program called Trainable Weka Segmentation. By “training” the program to identify cells, specifically where they start/end, the program co...

We started off our week with a congratulatory acai bowl trip to celebrate Preston’s acceptance into a training grant program. Acai bowls in California top Playa Bowls (no question about it). From what I can tell, its a pretty huge honor to be recognized by this grant, but he’s really humble about it. On Monday, Preston and I decided that testing antibodies that have never been tested on prostate epithelial cells before would be a good objective for my first Western blot on my own. We needed to probe for ASCT2, a glutamine transporter, and GLS in order to determine if their corresponding antibodies are functional. Antibodies are crucial for Western blots because they bind to the protein of interest (POI), allowing for us to qualify its expression after imaging. As such, Preston wanted to make sure they worked by probing for ASCT2 and GLS on three different cell lines. Cell lines are commercially purchased human cells that have been immortalized (modified to grow indefinitely) by telome...

I woke up early on Monday, June 11 th in preparation for any unforeseen events on my way to the Fuccillo lab for the 1 st time. Thankfully, I did – I got lost and turned a 15 minute walk into an unfortunate 40 minute walk through Philadelphia, in particular Penn Campus and CHOP. In hindsight, trying to find a shortcut on the first day may not have been the best idea. To add, Apple maps led me to wrong building, but Google maps saved the day. In the end, however, it all worked out as I arrived just in time to meet with the postdoc I would be under, Dr. Kyuhyun Choi, or Kyu for short. He led me to Dr. Fuccillo’s office (the PI) where we discussed some plans for the summer. I was then introduced to some of other the lab members, which included 2 undergraduates, 3 graduate students, a lab manager, and 1 other postdoc. Everyone was warm and welcoming. Kyu also toured me around the 3 lab spaces the Fuccillo lab uses. These include the main lab area, a separate testing area, as well a...

I’m about to wrap up my first week in the lab and I learned that Californians do in fact use the word “gnarly” as much as one would think. The graduate student's paper! View from the lab My lab is located on the UCLA campus right next to Westwood, which is a great spot to grab food or go shopping (basically I’m in my happy place). And everything here in Los Angeles is vegan! Even my PI! I was a little bit worried that I’d be living off Clif Bars for lunch, but the students in my lab are so welcoming and friendly that I have been blowing through my allowance easily by going out to eat almost twice a day, everyday so far. I feel so at home in my lab--the people are amazing, the location is great (and sunny), and the science is incredible. Before starting my internship I was a little concerned that I would be placed in a corner to pipette from 9-5, but I got more than I could ever hope for. The graduate student whom I am shadowing, Preston, explains everything in so...