Hello and welcome back to my blog! Thank you so much for your willingness to keep up with my discoveries each week. This week, in particular, I’ve learned a great deal! The George Lab at UCSD has undoubtedly become an experience I will never forget, and I’m grateful to be working here!
Mon. June 30, Day 22-
Perseverance was the big lesson of today. I got to spend my day with Max, Elizabeth, Sonja, and, of course, the rats! Max is an undergraduate student and has started his master’s project. This project is dealing with ethanol vapor, a little different from the cocaine and oxycodone that these rats are usually put under. Today was to test if the chambers would work for his future
1 set of chambers and the computer that runs the program
experiments. They told me that they have been testing these chambers honestly since April and haven’t reached their desired level until now. My first week here, I remember Sonja and Max testing out these chambers, and they unfortunately didn’t work yet. Elizabeth worked with multiple people (Olivier, our PI, Giordano, a previous scientist for the George Lab, and Morrie, the person who made the chambers) to fix the problems, and they were able to accomplish it! So today was all about testing the new system to see if everything was working out. Before putting the rats in, we had to test to see if the breathalyzer was working well, and we recorded the amount of ethanol vapor in each chamber. Max wanted to make sure his protocol was going to work, so he handed me the instructions to measure with the breathalyzer, and had me follow
Breathalyzer and procedure!
his procedure! Along the way, Sonja and Elizabeth helped make a few tweaks, but in the end I did get it to work! Was it necessarily right? Not quite, but the way Max did it was consistent, so it was okay that there was a few outliers. Once recording the initial amount of vapor in each chamber, we put in all of Max’s pilot rats into the chambers and pushed out vapor for an hour straight. Although the rats won’t be getting alcohol vapor straight pushed into the chambers for the actual experiment, this was to test that the rats would take in the gas and get drunk, basically. The alcohol is a vapor because the rats won’t drink that much alcohol in one sitting. The actual tests are supposed to be 8 hours long, and it’s similar to GWAS. Instead of a button to push, the rat get a little nose poke to get a puff of ethanol. After the rats were in there for 30min, we went in to check that the rats were in fact still alive. We noticed that some of the chambers started building condensation which wasn’t necessarily a good thing. When Sonja came back into the room, she noticed that two of the tubes had liquid ethanol in them instead of the gas it’s supposed to be. My hypothesis was, was that the tubing was too cold to keep the ethanol in the state it was supposed to be, and Sonja confirmed it! She said that the insulation of these tubes weren’t as good as other tubes, so we would have to go and replace them. The rats though, were taken out after an hour of ethanol, and you could tell the little guys were a bit tipsy! We put them on the surgery table to poke their tail to get blood for tomorrow, and some of them kept wanting to run off of the table because of how doozy they were. It was a little funny to watch. After all the blood was collected, Sonja and I headed to Skaggs to do IHC Day 1 on the remaining slides that haven’t been stained yet. It was truly a great day of learning, and it was so cool to be able to witness it all!
Tues. July 1, Day 23-
Another day at Skaggs is always a great day in the lab! It was a pretty chill day today, but I got to hang out with someone who I haven’t necessarily connected with yet. Sonja and I started the morning with Lab Meeting, which hasn’t happened in the last few weeks because it kept getting cancelled, but it was fun to actually participate in the meeting because I actually knew what most everyone was talking about! Compared to my first Lab Meeting to now, I think I’ve definitely grown, from my relationships to people in the lab, to developing understandings of science and how the lab actually works. After the meeting, Sonja and I completed IHC Day 2 (which I didn’t mess up this time!), and bounced back and forth between IHC and imaging. When IHC was completed, Sonja headed home so she could work from there, to focus on working on her writing for grants, emails, and much more. Joseph also happened to be in Skaggs today, so when Sonja left, there was nothing better to do than learn from Joseph! Joseph is our tech at the George Lab who does Brain Clearing. Brain Clearing is exactly what it sounds like: clearing the brain! It’s this whole washing and bleaching process to get rid of it’s muscle, so the only thing that’s left is the tissue. Joseph explained that the protocol was similar to IHC, but instead of the protocol only calling for two days, Brain Clearing takes basically about a whole month to get to the brain you want.
How the brain looks at first vs…
Clear Brain!
I got to shadow Joseph as he was doing one of many clearing washes in the protocol. A lot of the procedure calls for washing and waiting, similar to IHC, but instead of washing the slices in PBS, Joseph used a clearing reagent called DCM to make the whole brain clearer. After that, I helped him label some tubes, and it was honestly a great time! I had a lot of fun learning from Joseph, not only because I got to learn about something I didn’t know about, but also because I got to know Joseph a little better! Although I’ve seen Joseph around Skaggs and sometimes MTF, I haven’t gotten the opportunity to actually learn about what he does or who he was honestly, so being able to get the opportunity to do both was awesome.
Weds. July 2, Day 24-
I had another fun day working on Max’s project with Elizabeth and Sonja! Today, we had an early morning appointment with
Here’s what the Analox looks like
what’s called the Analox, and it tests to see how much alcohol is in the blood. The protocol for the Analox is very specific, so we made sure to read it over and then reread it before we even started to turn it on. The Analox was a tool of Giordano’s Lab, and since we get to share spaces and equipment with them, instead of being in the basement of MTF, we got to be on the 4th floor- very exciting news if you ask me! The Analox goes through this system where 5 microliters of blood run through the tubes of the machine, and they determine how much alcohol in mg/dl is in the blood of each rat. A pattern I’ve noticed in a lot of machines in science is that the preparation and the calibration of them all takes longer than the actual running of the data. This is really important, though, as we don’t want to waste the material we’ve gathered to actually make the data we want, so making sure that systems work is really key to success. When we were finished with the Analox, Sonja went to work from home so she could focus on her research. Direct quotes from her, “I just need 4 hours where I need to lock in to finish this,” so that’s exactly what she did! Her lock-in session gave me another half day in the lab. I hung around MTF, back in the office with Max and Elizabeth. They finalized some findings for Max’s lab, and then Elizabeth headed to the store to get better tubing for the chambers. Max invited me to go handle rats with him, so of course I accepted. Although I didn’t actually hold the rats, I did get to see them being held in
Max being my model using the Analox
Max’s hands. The rats need to be handled at least three times a week so they get used to being around humans. They also develop a connection with their handler so they know that they’re okay, which I think is so adorable! I will say, these rats have grown on me, and I really enjoy working with them!
Thurs. July 3, Day 25-
Today was a really chill way to end of the work week. In the morning, I worked with Sonja in MTF for a bit. We went and checked the rats we did surgery on last week, and they were looking better than they had been. This week, when Sonja checked on the rats, one of the rats’ surgery sites kept getting infected, and once that happened, the rat kept pulling it open, so Sonja kept on having to glue the cut back together, and then yesterday, she sutured the cut closed, so looking at the cut today and nothing being there was great news for us. Before checking the rats, though, Sonja was telling me about a different project she was working on, and it was quite a toughie. I got a great biology lesson from it though, and it was really interesting to learn about! She was doing a huge problem on DNA sequencing, trying to find the gene of interest in the coding portion of the genome, rather than the whole genome itself. In the coding region, she highlighted specific nucleotides to show mutations to look out for when using the region, as well as the gene of interest we wanted. It was quite difficult because DNA is completely different from one sequence to the next, so in each genome we were looking in, we had to make sure we found the right set of nucleotides to go along with it. Once we had one sequence figured out, we headed to Skaggs for the afternoon, where we found Natalie imaging some slides. Sonja continued figuring out the other sequence she needed, while I shadowed Natalie. A lot of the slides we took pictures of today weren’t as good as previous ones, but we still took pictures of them because there was some expression. The last slide, though, was beautiful! Natalie and I were taken aback on how good the picture looked, and were so excited to finally get a good picture. Classifying
Natalie and I imaging!
“good” took a little bit for me to understand, but you really want to see the virus express in the injection site, as well as the virus surrounding the cells. There shouldn’t be too much virus, but enough to show that there is something going on there. The stain being used is called DAPI, and it shows up blue on the computer. The virus expression, aka mCherry, is red on the computer. You should also be able to see the axons and dendrites kind of moving from cell to cell, surrounding each one it affects. Axons and dendrites are a bit hard to see, but axons are kind of like the tail of the neuron body, and dendrites remind me of the neuron’s fingers. The axon’s job is to send messages to other neurons, while the dendrites grab the message from other neurons to interpret.
Fri. July 4, Day 26-
Fourth of July Friday! It was so fun to have yet another holiday to celebrate with these amazing people! By now, you should know where we went on our day off; the beach, of course! We had a great morning and afternoon at Mission Beach, enjoying the sun, reading our books, and even making bracelets! Our host mom, Jennie, had also brought boogie boards with her, so we took them to the beach with us. I had never gone boogie boarding, so it was so exciting getting to learn how to ride it! Once I finally got the hang of it, Azari and I decided to catch a big wave together. Very big mistake. Unfortunately, the wave ate us up and we were quite literally water-boarded. Thank goodness the family we ran over was okay, but I still feel really bad. Azari and I gave up on the water and started building a sand castle with Maiya while Lana and Shiloh kept boogie boarding. After the beach, we went home to freshen up and
Azari and I channeling our inner dads grilling the burgers!
dress up festively for the night! Azari and I got to grill burgers, while Lana, Maiya, Shiloh, and Jennie were in the kitchen making toppings, salad, and fries. During dinner, we played a game of 5th Grade Trivia which really humbled my knowledge about everything in the world, however, we moved onto making s’mores once the darkness started to hit the backyard. Our backyard had a great view of the fireworks, so we got to eat our s’mores while watching the beautiful fireworks light up the night sky.
Sat. July 5, Day 27-
My uncle and my brother were able to visit me while I was here in San Diego, so I got to spend the beautiful day with them! We spent the day at Sea World, where I got to meet so many different underwater creatures. I really enjoyed the time I spent at the park, as well as with my family. It was a great addition to this holiday weekend! I think my favorite animal I saw there were the orcas. They were so smart and performed amazingly in the show they had. I also enjoyed watching the jellyfish float around, and they were quite breathtaking. Although it was a little different from spending time with the girls, I still had a great time!
Jellyfish at SeaWorld w my uncle and brother!
Sun. July 6, Day 28-
I got to spend another day out in San Diego with my uncle and brother before they leave tomorrow. We went to the San Diego zoo, and the animals were so cool! My favorite thing we did at the zoo was watch this 4D movie about the different animal groups. Afterwards, we went and had dinner with my cousin-ish and her family! She’s not actually my cousin by blood, but our families are close enough that we consider ourselves one family. The last time I saw her and her family, I was pretty young, maybe 6 or 7 years old, so it was great to catch up after all of these years.
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