Science Project

Here you will find my main project. Back in 2020, I got invested with the increasing rates of vaping in youths. In my high school, too many teenagers were addicted to this electronic nicotine delivery system (ENDS). This not only had great impacts on their physical health but also on their mental health and their school performances. Arriving at Collegial international Sainte-Anne, I realized that the rates were probably worse in this new environment. The Honours Science program, gave me the opportunity to get involved in this subject to, maybe, find a way to influence a couple vapers to change their bad habits.

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Review of Litterature

Science Conference

Preparation and Background Work

Right before starting my experiment or even before formulating my question, I had to read the latest news concerning my subject. I went through many papers, read "Big Vape: The Incendiary Rise of JUUL" by Jamie Ducharme and listened to the famous podcast "The Vaping Fix" by Wondery. I also attended the E-cigarette Summit USA 2021. For more details visit my Review of Literature page as well as my Science Conference page.

Question, Protocol and Materials

I established the following question for my experiment: What is the toxicity level of e-liquids for c. elegance and drosophila when axing on the concentrations of menthol, THC and propylene glycol?

To realize this experiment I will first analyze the components of three different e-liquids; a mint flavored one, one without nicotine and lastly one that contains THC. Afterwards, during three experiments, I will inject different concentrations of e-liquids in the drosophila and c. elegance environment to determine their toxicity levels.

After finishing my protocol, all I had to do was to buy all the material I could be needing for this fantastic adventure.

Protocol

Bioethics Statement

Since I will be working on living creatures, my project needed to be approved by a bioethics committee. I passed the last few weeks thinking about the moral side concerning my experience. What would the value of this project be compared to the loss of the Drosophila and C. Elegans? Could I have used AI technologies to obtain the same results? What measures will be used to minimize the sufferings of the living beings?

These types of questioned guided the writing of this statement.

A Wind of Change

My project has grown quite a bit since. Because of that, it was decided to split it in half. The first part will be the cell experiment and the toxicology background. The second one, probably done next year, will be the one done on living beings. The protocols have been updated and the list of materials have been approved. I am very excited to be conducting this first part in May 2023. The adventure begins.

A Long-Awaited Begining

Right before the end of my semester, I had the chance to finally start the first part of my experiment. From May 25th to the 3rd of June, I was mostly in the Sainte-Anne's laboratories. I grew as a scientist and developed not only my skills but also my critical thinking. I loved every second of it and I hope to be continuing this project next year. Here are some pictures of my lab adventures :)

First Day in the Lab!

I started my experiment with the realization of an ELISA test. The goal was to determine the approximate concentration of vitamin E acetate in e-liquids which had been heated up to 180 Celsius.

And Now We Wait...

ELISA protocols require patience, since the samples and the added reagents need to incubate at certain temperatures for certain amounts of time.

New Techniques

Here I am with the multichannel pipette!

Some Textbook Results

Here is my ELISA 96 well plate with the final results. I am very happy to have gotten conclusive results. As you can see on this picture, the kit with which the ELISA test was conducted used reverse absorption.

Complications

When the THC sample was heated up its container melted! I succeeded in saving just enough to do the ELISA test but had to go and grab some more for the rest of the experiment.

A Break in Equipment

Because of technical issues, Tania, the lab technician, and I resorted to spectrophotometry for the THC sample. We converted the resulted we had gotten at Ahunstic's labs to make an equivalence between the two techniques.

A Notch on my Belt

In the end, I really loved this experience. It thought me the uncertainties of science and perseverance. I was very proud of myself as I know ELISA test are very sensitive and can fail for many reasons. Science is really my beloved field.

Loving the Little Things

As I was learning to work with the cell cultures, I fell in love with the beauty of it all. The microscope can really reveal beautiful natural patterns that I never would've thought possible.

Preparations

Here you can see my first dilution of the SF9 strain. The samples sent by ThermoFisher were very concentrated and I needed to figure out how many milliliters were needed per petri dish for them to continue thriving.

New tools

Here is the first ever picture I took of the hemacytometer. You can tell by the fact that its not centered properly. Every tiny dark blue perfectly circular dots are viable cells. With bigger magnification you can see them more clearly.

SF9 Family Picture

Here are all my SF9 cultures. They were "dancing" on the orbital shaker day in day out. I placed them in the incubator to make sure their temperature stayed around 27 degrees Celsius.

At Work

Working under the laminar flow cabinet was a really challenging experience. I had to think about all my movements to make sure there was no contaminations.

My Very Own Cells

The buccal cells were collected from my own mouth. I had to be extremely careful about contaminations since many bacteria live in the buccal cavity. I wouldn't have wanted them to get infected!

1, 2, 3...

A different counting technique was used for the buccal cells. It was more tedious as I had to count them all by hand. As per usual, I was amazed by the beauty of it all. Here, the pointer shows a tiny buccal cell.

Setting Up

For 3 consecutive days, I was vaporizing the e-liquid for it to come in contact with the cells. The machine you see is a custom made sous-vide machine. The change in pressure helped to transfer the vapor from one container to the other.

Unhappy SF9

Every group reacted in a negative way of some sort. Here you can see the deformities of the SF9. After being exposed, they started accumulating together and their shape changed. Their concentrations also dropped.

Unknown Substances

In other petri dishes, there were some unknown debris. My supervisor confirmed me that they were too big to be bacteria and were probably little accumulations of dead cells. Although we cannot be 100% sure.

Unhappy Buccal Cells

Next to the air bubble (the yellow circle) you can see an accumulation of cells. A theory would be that the buccal cells exploded and the DNA bundle agglomerated its surroundings creating this dark patch.

Contamination

As much as I tried to be "perfect" under the laminar flow chamber, some mistakes were made. Two groups were contaminated with what is believed to be Candida Albicans, a common fungus found on the human skin.

Fungus Hair

After doing a biopsy of the fungus substance that appeared overnight in my samples, I looked at it closely under the microscope. Here is what I saw!

May this Adventure Continue!

I am very grateful to have had this opportunity as it really made me grow as a scientist. I hope to never lose this passion I have for science!