As a part of trying to characterize the cells that I have been studying and working on, one of the techniques that I had to learn was called Flow Cytometry. It is a technique where each cell is counted for and can be characterized on all physical and surface chemical characteristics – by this I mean the shape, size, contour, different stains etc. Before running the sample to the Flow machine, you have to prepare the cells with incredible precision and accuracy. There are many steps in which this can go wrong and hence, for the first two times, I was just shadowing my mentor. The third time, I was doing everything on my own with my mentor watching on. However, the fourth time, I was all on my own, trying to nail this complicated, time sensitive 3 hour long procedure all on my own.
Diagram showing the machine and its basic workings.
I read up on my notes and had them handy with me at all times cause I did not remember every small step. I had to run 10 samples all at once, so doing that for 10 different cells and markers was going to be hard. I started off strong, following the protocol and abiding by the time frame. By the time, the one hour incubation had arrived, I had a good feeling about the experiment. One hour later, I started with the next steps. I was feeling good until I suctioned off the cells from the side of one tube by mistake. It was truly heartbreaking as I would not get much data from one of the ten samples and that could hamper my experiment. Nonetheless, I continued with the rest. As I ran it through the machine, that one sample had very skewed graphs because of the low number of cells. I was completely shattered at the point and was scared to tell my mentor that I messed up after all the confidence she put in me. She reassured me saying its fine, it is my first time, we all make mistakes.
I was yet left disappointed in myself but wanted to prove that I have what it takes to do it successfully and that I can learn from my mistakes. The week after when we had to do the fifth run, I did it all on my own and got the results we were looking for which validated previous data.
I was so happy that I could overcome my failure and that gave me the confidence to do everything else on my own after learning the techniques. In addition, it provides a very useful experience for me and one that I can learn from!