The first few days working with dii staining I had a few issues with the entire worm being fluorescent making it impossible to actually see the amphid neurons. This was just caused by the dye clumping at the bottom of the centrifuge tube and was easily solved by making sure I mixed the solution better. The other difficulty came with identifying exactly which neuron was which. This was difficult because almost every worm I looked at was oriented differently on the slide and sometimes all the neurons looked too clustered to accurately identify.
I have looked at a decent amount of these worms now and from what I’ve observed there is not a difference in expression between males and hermaphrodites.
I have also still been working on getting a successful cross between my him-5:dmd-6::GFP and him-5:AWA::mCherry worms. I’ve tried numerous times and still haven’t had any success. I’m observing a few more crosses today that I set up so hopefully I’ll have some luck.
Things have been going pretty well the past two weeks! I’ve been scoring my prab-3::dmd-6 worms as well as my ptdc-1::dmd-6 worms. Just yesterday I learned the technique of dii staining. The actual process of the dii staining is very easy; you essentially just let the worms soak in fluorescent dye that is mixed with water. Then after an hour and a half or so you pipette that liquid dye containing the worms onto a clean plate. The more difficult part is actually looking at and identifying the stained cells on a microscope. It stains the amphid neurons in the worm and some of these neurons are clustered, so depending on what angle you’re looking at the worm from it can be challenging to tell exactly what each neuron is. Once I observe more worms in the next few days though I’m sure it’ll become significantly easier to identify these neurons!
This past week I began scoring the prab-3::dmd-6 worms that were injected last week for odr-10 expression. Once my ptdc-1 worms are grown out, most likely this upcoming Monday, I will begin to score those as well. It’s exciting to finally be getting some data and seeing this project move in the right direction.
I set up the cross that I wrote about last week and as it turns out it’ll be a little bit more difficult to determine whether or not it was successful. As of right now it seems as if it worked and I should be able to begin observing dmd-6 expression in AWA next week.
For my project this summer I will be looking at dmd genes and the how they effect expression of odr-10 in C. elegans. Previous research has shown that these genes play a role in the determining the sexual state in the nervous system, but a better understanding of it all is still needed. I will also be trying to confirm that dmd-6 is not expressed in AWA. Thus far I have been working on gateway cloning and initially had some struggles, but this past week I have been successful! In this upcoming week I am planning on setting up a genetic cross in order to determine if dmd-6 is being expressed in AWA.
These past two weeks alone I have already learned so many new skills and techniques that will be incredibly useful in the near future. I really look forward to seeing how my summer project progresses and keeping you guys updated weekly with all of my successes and (hopefully very few) failures!
My name is Connor Schroyer and I am a rising senior at Ithaca College in New York. I am a Biology major and Mathematics minor. This past semester I worked in Te-Wen Lo’s lab with C. elegans and I am extremely excited to have the opportunity this summer to continue expanding my knowledge and conducting research with this model organism. Through this summer research program I hope to even further understand C. elegans and learn skills and techniques that will be useful this upcoming school year in my individual research project and into the future when I’ll ideally move on to grad school. At Ithaca I am also a member of the Men’s Swim Team so a lot of my time outside of class or lab is spent at the pool. In the free time I do have, I enjoy going for hikes around Ithaca’s numerous State Park’s and attending concerts whenever possible.