This past weekend was a nice bit of fun before getting into work this week as me and a few other REU interns went up to Portland for the weekend. We went to the International Rose Test Garden, the Oregon Museum of Science and Industry, at least 5 thrift stores, and had some really good food along the way. This week I also conducted more trials of the boundary experiments with the larvae to see if they would actually move to interact with the beads rather than just stopping right after colliding with them. After remaking slides and sitting in front of a computer for a couple of hours just watching the larvae, I was able to catch a couple instances of them actually swimming and encountering the beads!! The best instances were when the larvae would run into the bead and it would run along the edge of the larvae’s surface so I could observe how the skin cells fluorescing were primarily those that the bead was directly touching. I guess it just took messing around with the coverslip that goes over the slide; the larvae might have been a little too squished last time (but they could still swim so I’m not exactly sure if this is true).
This week George also showed me how to tether the larvae using a really small pipette that can grab and hold the larvae so we can expose them to different stimuli. I also took a plankton tow, which is where I use a really small net to catch small organisms in the water and took them back to the lab to use them in my experiments with the tethered larvae. I specifically wanted bivalve veligers, which look like little clams, as they swim really fast. We haven’t yet conducted trials with the tethered larvae and the fast-passing swimmers, but we always have next week!!
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This past week has been fun both in and outside of work. Last weekend was super fun as me and a few of the other REU interns went to Eugene for the weekend! It was such a nice chance to get out and actually experience the summer heat I’m used to (lol). The transition from Georgia to Oregon has had me missing the heat of summer (crazy right?). We actually ended up visiting this really pretty watering hole, where there was rock sliding and some pretty cool places to jump off of too, I just wish all of the interns could’ve come along! This week I’ve been working more with George and Maureen to conduct more trials on my sea star larvae. Our main goal was to make a slide where they would interact with a boundary so we could image them under the fluorescing microscope to observe any signaling. We did run into a few difficulties as we needed something that we could add to the slide that would actually work as a boundary. We tried 10-micron (very, very small) glass beads (didn’t see much), considered sand (too big), and finally settled on these plastic beads that were about 100 microns (about ten times bigger than the glass beads). However, after finding these plastic beads, when we tried adding them to the slide, they just moved to the edge of the water droplet instead of actually sitting on the slide, not ideal when our experiments need the larvae to be able to collide with the beads. I tried supergluing them to a prepared slide; however, it was hard to separate each individual bead and the slide ended up being too thick to actually get any good videos. George helped by giving me the advice to treat the beads with a protein that would keep them from moving to the edge of the water droplet and clumping together, and it worked!! I’ve done some imaging of the larvae interacting with the artificial boundaries, but it’s really hard to observe localized signaling because for some reason the larvae just stop swimming?? Guess we’ll just have to keep on moving, and hope the larvae do to. Until next week… :)
This week has been a lot of lab work, but I can’t say I haven’t been enjoying it. We started this week imaging larvae with the GCaMP sensor for detection of electrical signaling. George and I even went down to the dock to take a plankton tow, which is when you cast a net with really small threading to catch microorganisms you wouldn’t typically see. Using some of these organisms that swim considerably faster than our larvae, we subjected the larvae to some fast-passing events to observe whether the larvae showed a response, i.e., lighting up under the fluorescing microscope. We also observed passing events of other larvae to detect electrical signaling events, and guess what, we did!! Not only did they do it once, but twice!
While working with the larvae, I’ve also been working on that second project I mentioned looking at how caffeine treatment can disrupt meiosis and cause sea star eggs to develop into cells that have twice as many chromosomes as they normally would. George and I microinjected sea star eggs with two fluorescing proteins, one that highlights the microtubules that make up the spindle that forms during meiosis, and another one that lights up the chromosomes that are split during meiosis. We then took matured sea star eggs and under a microscope, took a few videos of normal meiosis. You can see in the video below, the spindle apparatus (left panel, gold color) assembles, and microtubules attach to chromosomes (right panel, blue color) to bring them to the middle and then segregate into two cells (one is really small), after which the apparatus disassembles. When cells were subjected to caffeine treatment prior to the separation of chromosomes, we saw a complete disassembly of the spindle apparatus, preventing meiosis from continuing. However, when we flushed the caffeine from the eggs with sea water, they completely recovered and completed meiosis (I’ll add videos of this next week)! The recovery rate was really fast too!!
Socially, this week has been pretty chill. I’ve taken a few trips to OIMB beach, helped cook a really good meal with friends, and taken a few walks through the woods. Being able to be in an environment where I can both grow academically while not sacrificing these small experiences has been really enlightening. I’ve found myself more excited to head to the lab with a clear head after a quiet walk wandering through the trees (maybe I just really like being in the forest lol). Well, until next week… :)
This past week at OIMB has been a straight shot into working on my project. Even though it doesn’t feel like I’ve done much in terms of fleshing out an entire designed experiment, I really value George’s approach to research. Being able to work together to both work hands-on and learn as I go with the understanding that science isn’t always going to give you what you expect has really encouraged me to ask questions and just be more enthusiastic overall about the work that I am doing.
Throughout the week I’ve been working with both George and Maureen to generate starfish eggs that have either a calcium sensor or added fluorescing proteins that essentially makes certain parts of the cell light up under a fluorescent microscope. Looking at those with the calcium sensor, we want to check that both the eggs and the larvae have the calcium sensor by looking for this fluorescence (lighting up). Since last week, we have fertilized and cultured larvae with this GCaMP calcium sensor to then study and image movies to detect signaling observed thanks to the calcium sensor. We even introduced different stimuli, such as adding food to the slide with larvae to see whether or not that would show any obvious response seen as the flashing of these skin cells.
Now, conducting research in the lab along with learning how to use at least 5 different microscopes has been fun, but I was super stoked about the opportunity to go camping this past Friday along tide pooling the next morning. I had never been camping or tide pooling, so this was a completely new experience for me. Sleeping outdoors wasn’t the most comfortable, but the experience let me know that I’m definitely down to go camping again. Tide-pooling that Saturday morning, while a little chilly, was great (that is if I disregard the muscle soreness I had after, I’m a lot more out of shape than I thought lol). I had never seen anything like it, trekking through water up to my waist, trapezing (aka trying not to fall) over algae covered rock (super slippery btw), and being able to observe just how these organisms we work with in the lab exist in the wild was something I never did a great job of imagining. But seeing these animals in their natural habitat only made me more excited to delve into the behaviors that can’t be observed in this environment but need to be seen up close and personal in the lab. It’s safe to say I don’t plan on accidentally slipping and falling on a sea urchin while tide pooling any time soon. So, until next week… :)
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