Nothing excites me more than coming into lab, and getting ready to dig my hands dirty conducting experiments. The familiar smell of ethanol and tight latex gloves squishing my hands sets the dawn of a new mission: flow cytometry. For my project, this is the technique that I will be using to conduct most of my experiments with. Just to give some background, flow cytometry is a method to analyze granularity and identify cell type. The mechanism behind it is that as the sample flows through the machine, each single cell passes through a laser, causing deflections in light which is analyzed to tell cell type.
Now that we are past the technical details, here are some lessons I learned from the past week while attempting to complete flow!
1. Math skills are still vital. Even in biology.
From what I remember from school, biology class was one of the sciences that required the least amount of math skills. And although I’d like to say since I picked a biology based research topic I could finally escape math, but alas it came back to haunt me. I was trying to do get 2 million cells from the original sample for flow, and after several trials of calculator punching, I finally came up with the right answer. I won’t get into the specifics of the calculations, but it ended up just being a simple proportional cross multiplying problem. It took me a solid 10 minutes. Lesson learned? Pay attention and remember your math skills!
2. In lab, everything should be handled with 10x caution.
The first time I did flow, with my mentor watching behind me, I could feel him wincing as I transitioned from each sample to the next. You see, the flow machine works by sucking in the sample through a tiny tube, and in-between samples, the machine just ends up sucking air. This could end up hurting the machine and distorting the data, so you have to be quick when changing the samples and prep for the next sample while it is still analyzing its current sample. I, unaware of the consequences, blissfully took my time. When I realized the mistake, it suddenly became an aggressive arm workout swinging samples left and right, spilling portions on the ground in the process.
It was understandable why my mentor winced. Grover, the Fortressa flow cytometry machine, is worth whopping $20,000 , and breaking it would be put lightly.. tragic. But like friendly and strong Super Grover of Sesame Street, it still stood strong and (hopefully) forgave my mistakes.
Grover in all its glory
3. My lab members are the most AWESOME human beings on earth.
After I ran my flow experiment, Grover connected to a computer, analyzes the data and puts them on graphs. To further analyze the data into conclusive results, we use a method called gating. In order to weed out the populations that were not necessary, we draw a “gate” or a rectangle/polygon using the FlowJo program, and create a graph focusing on that population. The process is quite obscure, and as I was trying to gate I felt like I was just randomly drawing shapes onto the graph. It was kind of confusing at first, but my lab members were there to sit behind me, and guide me through the process. To be frank, this goes for every aspect of the project, not only for flow. Sooo many times while I was doing other experiments, I had to stop every few minutes and make sure I was doing the right step. My lab-mates were super supportive in answering my endless questions, making me realize how really collaborative science research is.
My messy lab notes detailing the different types of gates
Not only did they make the process more understandable, but they make coming into lab even more fun! Weird jokes from reddit (science and non-science related), talks of 2020 election, and random fortnite dancing in the lab, constantly make me laugh and smile. They truly embrace the motto of work hard, play hard.
All in all, the first week has just been about understanding and practicing the method of flow cytometry. Next week’s blog will go further into explaining the overall project and creating growth curves!
-Till next time, Mabel