Week 4


I have finished the set of slides I was imaging! For the rest of the week I will be working on getting training and certification in order to work with the mice. The lab is going to be doing many behavioral tests on the mice in about four weeks, and I will be staying past the end of the senior project to help with that.

I put copies of the left hemisphere of two different mice brains that I imaged. The top one has lots of labeling (it looks like little green dots), which means it is probably a normal mouse that was sleep deprived, while the bottom one has no labeling, which means it is probably one of the knock out mice. I know the image is not very clear, but if you look closely on the original image you can see axons/dendrites coming off of the green dots of the neurons.

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Happy Friday!

I am got to run a gel again today for DNA testing. I attached a picture of the results below. The lab manager trained me on how to tell the genotypes based on the banding. Basically, the top band is only present if the mouse has the knockout trait (no functional Egr3 gene), the second band shows if the mouse has the Egr3 gene, the third band shows if the mouse has the X chromosome (they all do), and the fourth band shows if the mouse has a Y chromosome.

Just as a recap of Mendelian genetics, if the mouse has the knockout and normal Egr3 bands it means they are a heterozygote (written as +/-) and if they only have the knockout trait (-/-) or only have the normal wild type trait (+/+) they are homozygous. If the mouse has the trait, they will have a band at the first, second, third, or fourth place, and if they don't have the trait there won't be a band there.

 The genotypes of the first row from left to right are: Male +/+, Male -/-, Female +/-, Female -/-, Female +/+, Male +/-, Male +/-, Male +/-, Male +/-, Female +/-


I am also still doing the training to earn all the certifications needed to work in the vivarium (the room with the mice). It is a long process to get certified! I still have one more online training to do and I need to fill out a form about the vaccinations I have gotten and any allergies I have, so the lab can determine if it is safe for me to work with the mice.

In a few weeks, probably around week 10 of my senior project, the lab is planning to do behavioral tests on the mice, and I will be helping with that. I am also going to continue volunteering at the lab over the summer.

Week 3


I know it is just the beginning of the week, but I wanted to give an update because I did something different today. I got to play with dry ice! Just kidding, you can't play with it and you need to be careful with the dry ice because it is very cold and can burn you.

I was helping out someone in the lab who needed to access certain mouse brain slides from the freezer. Basically when they sliced up the mice brains they mounted all the slices on slides and now they found out what regions of the brain they specifically need to look at so they are accessing slides with those particular regions.

The slides need to be kept frozen, so we had to stand in the cold room (basically a small room that feels like you are standing in an industrial sized freezer) and leave the slides in their box sitting on dry ice so they stay cold. Then, the person I was working with would look at the tiny brain slices on the slide to figure out which ones had the regions they were looking for.

 It was a cool experience, but I had dressed for a typical Arizona summer, so I was freezing! Anyway I'll make more updates throughout the week.

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Hello again!

I wasn't able to do mouse brain imaging today because someone else is using the microscope, so instead I was taught how to make gels for DNA testing the mice. Basically with electrophoresis DNA samples are placed in wells on the gel, and then the gel is put in a tank with a current running through it. Smaller pierces of DNA are pulled through the gel more quickly than larger pieces, so at the end there are distinct bands of different DNA pieces in the gel. Looking at the banding pattern tells the lab whether the mouse is normal, a knockout mouse, or a heterozygous mouse, and it also confirms the sex of the mouse.

I attached a picture of the banding on the gel after it had been run, and the electrophoresis tank that the gel was put in. The bands aren't that differentiated because we didn't leave it in very long since it was just practice.

 

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Hope everyone is having a great Friday! 

I didn't get to work on imaging the mouse brain slides much this week because other people were using the microscope, but I did get to learn a lot of new things. Besides the gel electrophoresis experiment I mentioned above, I got to learn how to run the dishwasher for lab equipment, and I shadowed a lab employee who was getting training on how to test the lab for radiation. 

Have a nice weekend!

Week 2


Hope everyone is having a great Friday!

First of all, you might have noticed the title of my blog has changed. I haven't changed my project topic; I just wanted to refocus it on particular aspects of the research being done in the Gallitano lab. I also edited my Senior Research Project Proposal, which you can see by clicking on the Project Proposal link to the right.

I spent most of this week imaging the slices of mouse brains, which is cool because I am learning which mice types have more or less fluorescent markings (the gene knockout mice have virtually no markings because they are lacking the EGR3 gene that would cause them, while the normal wild type mice have lots of markings on their brains). However, it is still a lot of sitting in a small room in the dark starring at mouse brains, so I was happy that I got to do something different today.

Today I helped out with a new experiment the lab is running where they inject the brains of live mice with a virus that inserts genes that the lab hopes will help counteract the symptoms experienced by the knock out mice. If the mice are better at adapting to novel situations, are less aggressive, etc. it could show an interesting possibility for treatment (at least in mice). My job to help out with this was to pipette 10 microliters of virus solution into different tubes, and then put them briefly in the centrifuge. I had thought pipetting in general was a little stressful, but pipetting virus samples is even more nerve racking! Still it was a really interesting experience.

I also learned how to use the autoclave today. For those of you who don't know, an autoclave is a machine that heats up things to sterilize them, and the one in the lab uses steam. I was sterilizing some pipette tips that would later be used during genetic testing of the mice (they need to be sterile so no foreign DNA messes up the results). When you autoclave things you have to put a piece of special tape on them that changes color if it gets to the correct temperature.

Anyway, today was a fun end to the week!  I feel like I am learning a lot, not only about autoclaves, pipettes, and mouse brains, but also about what working in a lab is like. Thanks for reading!

Week 1


Hello! I just wanted to give an update on my project.

Everything has been going great in the lab. Yesterday I sat in on a meeting that summarized the research the lab has been doing recently. To give a very brief summary, the lab "knocked out" a specific gene in mice, and used these "knock out" mice as a model for schizophrenia. They found these knock out mice were hyper-aggressive and seemed to adjust poorly to novel environments. Now, they are trying to see if they can use viruses to change genes in the knock out mice in order to reduce their symptoms.

Today I got a tour of the lab, and safety training. I was also taught how to use their pipettes. Finally, I got to see a mouse brain get sliced into thin pieces that can then be mounted on a slide to be examined under a microscope. I never realized about how small mouse brains are; they are really tiny, but you can still make out the different lobes and features like the corpus callosum. These thin slices are made with a machine that cools the brain and then moves it up and down against a blade (kind of like a deli slicer for meat).

Anyway, I just wanted to give an update, and let you know I might change my project topic slightly this week based on what I am specifically doing in the lab.

Thanks for reading!
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Hope everyone is having a nice Friday! 

On Wednesday, I was trained on how to image the slices of mouse brain. Basically I am putting the slides with mouse brain slices under a microscope that shines a special light on it to make the spots that were stained fluoresce, then I am using a computer program to select each hemisphere of the brain so the camera attached to the microscope can take pictures of it. Or in other words I am sitting in a small room in the dark staring at pictures of mouse brains. I know that sounds incredibly boring, but it is actually really cool, because I can see lots of details, like the neurons that absorbed the fluorescent dye. I am taking these images at 10x, and at that power the neurons look like little green dots (everything is green because of the staining and fluorescent light), and sometimes you can even see the dendrites that look like tiny lines coming off them.

Enjoy your weekends!