This week has been full of adventures inside the lab and out! For the first 3 days of this week, Sherlyn and I were alone in lab, as both Professor George and Erin being out of town. Monday morning as I went to look at my slides under the confocal microscope, the fluorescent tagging didn't appear. I had to look at every step of the fixation and staining process to see where something could have gone wrong. Wednesday, I was able to fix the problem, as creating a new anti-body mixture using DM1A anti-mouse primary anti-body serum. Having the experience to trouble shoot and learn of my mistakes on my own has a great experience. It has given me the confidence to work on a lab alone, as even as things may not go perfect, I can figure it out and get back on track without needing a professor all time time. It makes me excited for the future, for when I am working in my own lab, creating my ow experiments. This week, I have been able to create document the sequence of events between two and four hours after meiosis two. In this time period, we are seeing the mitotic spindle form and separate the chromosomes in preparation of first cleavage. We see the cortical microtubules arranging on the surface of the cell as if to squeeze the cell causing first cleavage, but then tend to disappear. I am unsure as why that would occur and I am planning on continuing creating this sequence and timeline past four hours to hopefully understand the large picture instead of snapshots of what is occurring. Hopefully, by the time of poster presentations I will have a complete sequential order and hopefully a timeline to the DNA movement and microtubules order of the B. glandula embryo. For the adventures outside of the lab, Sherlyn and I went to the sand dunes just north of North Bend. Seeing two sand dunes, a beach, and swam in the North Fork of the Umpqua River. This REU program is about furthering your academic knowledge, but it also about experiencing a new area. It is amazing to be in Oregon for me as the environment here is incredible. You can stand on the top of a dune and look east to see the rolling hills and forest, then turn west and see the ocean. It was a breath taking landscape that I tried to capture in a photo, but it is never quite the same as seeing it.
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This past week has been much the same work so far. I am still working on creating a timeline from Meiosis 2 to the first few stages after the mitotic spindle forms. Of course, pro-nuclear fusion occurs between these two steps. I have had a set back this week, as two of my broods of embryos had trouble with the staining of microtubules. This upcoming week, I am going to replace the tubes of anti-bodies with new mixtures, as well as expirementing with differing anti-bodies. I have been using an anti-mouse anti-body, but we have ones such as anti-rat in the lab. They would stain for the same thing, microtubules, but would help us narrow down where the problem is occurring. When done correctly, we can see as above, the microtubules attaching to the condensed DNA, pulling half in each direction in preparation for the first cleavage. This is only at one specific depth in the embryo. With our confocal microscope, we are able to scan the embryo at many depths and the program we have puts all the images together. There we can see the embryo as a whole, instead of in slices. My plan is also to start staining very young, pre-meiosis 2, embryos to see of there is a consistent pattern in regard to where the sperm enters the cell. I Outside the lab this week, I have been able to go fly fishing a lot this past weekend at the local rivers. It was fun to catch a bunch of trout and get into the quiet of nature. I drove inland to the rivers, which got really hot really fast. I had to take a break mid-day to swim in the river. Even as it was ice-cold, it was very refreshing after being in the 90 degree sun.
Also, the REUs and the OIMB students got together for a fun relaxing evening at the OIMB beach down the street. I call it our "secret" beach because I didn't even know it existed until this week! It has been nice to grow our little community and get along with the students here. I can't believe it is almost half way through the summer! Concluding this week, I have started being able to do research on my own! I am confident and trusted enough to mate barnacles, extract their embryos, run an interval experiment, stain, and look at the embryos in the confocal microscope. WOW, that sounds like a lot when I write it out. Essentially, now I can create the images you see below. This is a barnacle embryo at Meiosis 2, when the polar body has already been extruded. On the left we are seeing the two pronuclei of the sperm and egg meeting! This is the where the Female DNA meets the Male DNA before the first division the cell will undergo. This is important to see as we are able to collect images in intervals, for example every 20 minutes, to watch for a general pattern at which the DNA is moving. We also watch the microtubules as they are the mechanism for how the DNA can position itself in the cell. The sperm initially send microtubules in all directions until it hits the female pronucleus(female DNA). Then pulls both sets of DNA together to combine. On the right, we are seeing the "mesh" of microtubules in the cell. This will help you understand about where the DNA are located in the cell. In the lab we observing these structures form and dissipate. Before the cell divides, we will see these microtubules start to break down and form new ones. Hopefully, I will be able to take a live video of these cells, watching the microtubules move as well as the DNA. Moving out of the lab, we were able to get on the OIMB boat this past Saturday morning. We trawled the bottom of the ocean about 150-170 feet down. Being able to see sponges and small bottom/near bottom creatures was very exciting! There were over 7 different star-fish species, decorator crabs, hermit crabs, many sponges, shrimp, jellyfish, sea cucumbers, and much more! Plus, it was a beautiful and clam day on the ocean, which makes the experience so much better.
Happy Fourth of July everyone! Our research has begun to build a vision for the rest of the summer! Over the first week we learned a lot about general and barnacle embryology. Having enough knowledge now, I have created my question that I will be focusing on for the rest of the summer. Is there a pattern that arises in regards to pro-nuclear meeting as well as where does the sperm enter the egg? To further explain, after a sperm and egg meet, the DNA of both must combine to create the young barnacle embryo. I am trying to learn about how these two groups of DNA find each other within the cell. We have and will continue fluorescent tagging these young embryos to see the DNA and microtubules and look at them under a confocal microscope. The microscope combined with the staining allows us to find and see specific parts of the embryos at specific depths through the embryo. By collecting a large amount of images from post fertilization to the first division the cell creates, we can analyze the images and describe what patterns are apparent and why they are doing what they do. Outside of the lab, we have been learning a lot about the local marine community by visiting the tide pools at low tide! As it is always fun to see the species that everyone knows, such as the starfish and sea urchins, it is outstanding how much life you would step right over without ever knowing its there. Limpets, Gum Boot Chiton, and Leather Back Chiton just to state a few new species I met. This cove have recently been overrun by sea urchins! The sea star populations have decreased due to the sea star wasting disease. The sea stars are a predator of the urchins, so a lack of sea stars have lead to a booming population of urchins. That's why you can see such a large field of purple urchins in this cove. In my free time this week, I have been visiting the beaches near Coos Bay. Especially towards sunset when the weather is nice, I enjoy sitting down near the water and either playing my guitar or watching the waves. I try to go to differing beaches every time, but these two spots are my favorite to revisit. For the weekend of the fourth, I was lucky enough to go home back to Illinois and see my family. Many of my cousins, aunts, and uncles came from out of town for a nice relaxing weekend. We were able to have a barbeque and celebrate my grandfather's birthday! I am excited to be back now and ready to continue trying to answer the questions we have regarding barnacle embryology.
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AuthorHello I am Gina Magro, currently attending University of Wisconsin-Stevens Point. I am studying Biology with a minor in French as a senior this fall. Archives
August 2022
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