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Visualizing Harmonic Convergence in Mosquito Mating

Wow, what a summer!!! I have some exciting news to report…I didn’t get bit by ONE mosquito all summer!!! Just kidding, my project is a little more exciting than that! I did it! I successfully put together and executed a project that I was a little iffy about back in May, and developed a new-found love for mosquitoes [fake news, don’t tell them I said that!]. I now like to be referred to as the mosquito whisperer, so if you see me on the streets, I will not respond to any other name.

But now, let’s get to the good stuff! Last time you heard from me, I was getting ready to start recording male/female pairs of mosquitoes. Now, I have about 7,000 audio and video recordings of these interactions, and I couldn’t be happier with the data I collected! The goal for this stage of my research was to observe whether or not mosquitoes actually communicate with one another to signal their interest in mating, or basically flirt. Below are the visual results of this from the previous study.

For my own recordings, I was able to detect the presence of these interactions by importing my audio files into a computer program called Audacity. Within this program, I could convert the sound file into a spectrogram that was able to clearly show me the frequencies produced by the mosquitoes in the recording. What the heck am I talking about, you ask?? Below is one example of a recording spectrogram that revealed a converging interaction!

But before I get into explaining the scary pink and blue stuff above, let’s talk about how I got these recordings in the first place- that’s the fun part (minus the 500 times mosquitoes got loose in the lab and attacked all of my friends…losers)! About midway through the summer, I changed some of my methods to make my procedure a little easier and reduce the number of casualties caused from pinning my little friends onto insect pins…yeah, they were not happy with me when they woke up from their nap to find themselves stuck to a wire…but, you got to do what you got to do for science!!!!! At the beginning of the summer, I was using insect wax (a yummy combination of beeswax and rosin) to fix these guys to their new home, but it turned out that the wax wasn’t strong enough to keep the mosquitoes in place when they woke up, and more often than not, they flew right off of the pin and straight for my face. So, I decided to try pinning them with a tiny amount of superglue, and it worked magically! The trick was to touch the super glued side of the pin to the mosquito’s thorax (pictured below) instead of their abdomen, which is where I was attempting to pin them when I was using the insect wax. When I tried to pin their abdomen with superglue, sometimes their wings would get stuck to the pin, making it a little bit difficult to get a good recording when their wings couldn’t move… Instead, their thorax provided a perfect amount of surface area for the pin without interfering with their antennae or wings at all.

Once I adapted this method, pinning them was a breeze! I kid you not, I could probably pin 20 mosquitoes within 30 seconds. You’re impressed, I know, I was too…Below are a few examples of my mad skills.

      

Don’t they look so comfortable and happy!? Next, I set up my recording stands, which were actually 3D printed ‘micro-manipulators’ designed by Backyard Brains! My company is so cool… These stands were used to fix the mosquitoes, with the help of some silly putty, for the duration of the experiment. They were perfect.

 

Now I was ready to record!! Below is a beautiful video of one of my experiments (I’m a little proud of myself, can you tell?) Make sure you turn on your sound!!

 

How creepy is that??? These noises will be burned into my brain for the rest of my life! But isn’t it also super cool? You can definitely hear the difference in sound between the two sexes, but can you hear when they begin converging?? Listen again.

If you’re thinking that it happens roughly 20 seconds into the video, lasting about 15, you’re right!! But just to be safe and make sure that the noises we were hearing were indeed interactions, I imported both files into MATLAB for a closer look

Here you can see the two different frequencies of the female and male (though there is a bit of noise blocking the females’ fundamental frequency). The key to detecting an interaction is to look at the higher frequencies, up in the harmonics, around 1200 Hz because this is where convergence will normally occur. And lucky for us, it did! On camera!  I was so excited I just about packed up and called it a day, but I really wanted to see some more interactions, so I pinned 8 million more mosquitoes and got down to business! In the end, I was able to successfully record, both audio and video, 49 male/female pairs, observing interactions in 33 of them! That means, in the small sample size I had, the pairs would communicate a love interest to one another 67% of the time! Gross, get a room!!!!

Nearing the end of my time in Ann Arbor, I finally finished recording, throwing in the towel for my beloved new hobby, and I was ready to start processing my data in the hope of making it a little more ‘Hollywood’ as Greg would say! Little did I know, this process wasn’t as appealing as I first thought, and on multiple occasions I considered playing with some more mosquitoes just to get away from the madness known as MATLAB. Lucky for me, I had a MATLAB expert living with me (Hmmm…maybe that’s why we became best friends since she couldn’t escape me anytime I opened my computer to work!) Christy helped me create the most magical, color coded, satisfying and all around perfect video of not only my little buddies interacting, but also a spectrogram underneath it that played in perfect sync with the original video recording! Brace yourselves…you will never see anything more beautiful in your life…

 

 

If you caught yourself replaying it multiple times, don’t fret, as you will catch me playing it periodically throughout the day just for fun. I’m not a nerd. But look, I was successful!!!

We also presented our research at a poster symposium at University of Michigan!

So now is about the time where we wrap up!!! Ah don’t make me leave!!!! But I am so happy with the work I produced this summer and I feel so lucky that I got the chance to be part of this program. Greg Gage, you are the best boss I have ever had (don’t tell that to my dad since he’s the only other boss I’ve had…) and I will be forever thankful for the impact you had on my life as not only a researcher but also an individual. I love you and your family to pieces, especially your little ones that taught me all about Peppa Pig, and are still convinced my name is ‘Dirt’. Wonder where they got that…cough, cough, Christy. I already miss you guys, and I haven’t even left Ann Arbor yet! I’d also like to thank all of the staff at Backyard Brains (Stanislav, Zorica, Will, Zach, Caty, Catherine and John), who made my time here so worthwhile and comfortable- I never felt alone even when my MATLAB would crash, or when my fellow interns would shun me for letting some mosquitoes loose in the lab…

And last but not least, thank you to all of the BYB interns that made this summer one for the books! You will all be a part of my life forever, and I can’t wait to see where our lives take us once we leave each other this evening. You’re all such wonderful people, and I couldn’t have asked for better friends. Love you guys!!

Backyard Brains forever!!!! (Tattoo idea, interns?????)


When Computers Hear the Birds Sing…

Hey there! Zach here with the Songbird Identification project for a quick update. Since the last post, I’ve been hard at work creating a prototype device to listen for and record songbirds. I began by creating a small circuit using a microphone and amplifier chip. This acts as a sound recorder and also includes circuitry to act as a trigger to start recording when sound is detected. Currently, the sound level can be set using an adjuster on the board and an LED light indicates when sound is detected. This worked, but I needed more power and a dedicated board before I could begin field-testing…

After proof of concept, I began work on the actual recording device using an Arduino M0 Pro microcontroller. I connected this to my circuit and an SD card reader and programmed the board to automatically record as a .wav file to the card when sound is detected. Having completed this initial prototype, I had the circuit turned into a PCB “shield” (an extender for the Arduino microcontroller). Once put together, the initial prototype boards looked like this:

At this point, it was about time to do a field test, so I took the board out into the woods near the Nichols Arboretum to see how well the board would pick up birds in the wild. Check out the video below to see it in action. When you hear the birds, watch the red LED on my board… it flickers as the birds sing! This is the visual indicator that the board hears the bird and has started recording!

Bird Box in Action

The next step will be to develop a weatherproof housing for the board. This is an important step for two reasons. First, I need longer term test recordings to make sure that my hardware isn’t running into any issues. Second, the housing itself is a key element to this project, as the end goal is to deploy these boxes for days, maybe even weeks, at a time. Be on the lookout for an update once I have my enclosure built!

My 4th of July Parade Hat, Ft. Acrylic, Laser cut songbirds!

Thanks for following the project thus far and stay tuned for more!


Recording the Behavior of Squid Hatchlings

Introduction to the project

Hi, everyone! Last week marked the halfway point of my time as a fellow here at Backyard Brains!  Recently, I’ve succeeded in building a rig and recording video footage of my Squid Hatchlings! I’m excited because it means I can start gathering quantifiable data! The squiddos have kept me pretty busy during these weeks, and they have been so fascinating to study (and photograph obsessively, like the proud parent that I am).

The main goal of my summer project is to document and begin to understand the behavior of squid hatchlings. Mostly I have been investigating how they complete the complex task of navigating away from where they’re born to seek prey and space to grow up. From previous studies, we suspect that they use their vision to help them navigate because they tend to swim towards sources of light, especially when they are very young. Since we already know a lot about what kind of light exists at the different depths, being able to specifically quantify the strength of their preference for different colors and intensities of light would help us understand where they like to be in the ocean at different ages.

In order to study how their light preferences change over time, I first had to figure out how to separate the eggs into individual containers that could still be aerated from a single pump. The containers needed to be able to let in oxygenated water, but keep hatchlings inside once they were born. The solution was a series of seven water bottle covered in tiny pinholes which rest in a bucket of water. As long as I check them regularly, I always know the age of a hatchling in any given bottle.

Home sweet home.

Next, I needed to write the computer software that could track the squid when I filmed them reacting to different environmental conditions. Since my camera and the background of the tank are stationary when I film, the solution was pretty simple. I break down the video into frames and find the average image over time to figure out what the background looks like. Then, I remove the background from each frame, apply some filters and am left with an image of just the squid! This method is called foreground detection and it is a commonly used method in image processing and computer vision. Once I have this simplified image, I can calculate the population of squid that are in each segment of their tank and see how they move over time. (Click the Gif’s to see the sweet action!)

 

Next, I needed to build an experimental setup in order to actually test the squid with variable conditions. After several failed attempts, I ended up with an acrylic box covered in black paper that has a hole for the iPhone to take a video through. This way, the only light that the squid are exposed to while I record them is the light that I produce and control. At first, I wrongly assumed that I should be testing squid with light from the top (since the sun is above them in their natural environment), and was getting very… boring results. Besides their sense of light, the squid also have a sense of gravity and are negatively geotactic, meaning that they like to stay at the surface of the water. So the squid in the dark would be at the surface of the water and I’d turn on a light and they’d just… stay at the surface. I wasn’t sure how to quantify this and I knew that these weren’t the results I wanted, since there was no differentiation of their reactions. I felt exactly like this:

My exact face after four hours of attempting to get results… I brought them outside the lab to see if the fresh air would help them? I just got soaked in the rain and still saw nothing interesting…

Finally, I had the idea to put the LEDs on the sides of the tank to test their reactions. I wouldn’t be forcing them to swim away from the surface, but I could hopefully still see them track the light. It worked! I could finally see the squid making an active effort to follow the light and can quantify their reactions to it.

Now I am in the process of designing the exact experiment that I want to study consistently across all groups of squid as they age. While I have been exploring and contemplating, I’ve seen some pretty cool effects. It seems that young squid (within 24 hours of hatching) react extremely strongly to low light levels (around 100 lux) and much less strongly to very high light levels (around 600 lux).

Two recordings from samples at <12 hours since hatching. The x-axis here represents the region of the tank where section 1 is the far left (in this case, closest to the ligt) and section 7 is the far right. The y-axis shows the average percentage of the population that can be found in the section over the course of a 2-minute video.

In testing some older squid, I’ve also observed that they don’t seem to react to red light. Perhaps they don’t see it at all, at the least they don’t care about it. It’s not conclusive yet, and I’d like to test more groups, but you can see the graph of their reaction below.

I’ve also seen that the older squid seem to react strongly to very strong intensities of light, especially compared to the younger squid. This might provide some interesting insight into the life of the squid and where they live in the ocean at different ages.

I think that that is all for now! After recording some more data I hope to have some awesome graphs to share with you that show how the squid react to low light, high light, and colored light as they age! With that information, hopefully we will be able to piece together the narrative of the early life of the loligo pealeii and improve our understanding of their abilities and behaviors.