I am sitting at my desk in the lab, still mad and frustrated that I can’t even pipette properly. My cell counter results are dismal. I don’t know why I am getting such poor percentages for cell viability – I should be getting at least 90%, not 47%. I followed what my supervisor did an hour ago: perform a serial dilution four times, pipette trypan blue into each of the four wells, then pipette a bit of sample into the cell counter plate.
Fortunately, my supervisor came into the cell room to check on me and helped me with pipetting for the cell counter step. He got 89%, close to the ideal threshold. “I don’t know what’s going on,” he said. I could sense that his voice was tinged with frustration, which I understood. I was also mad for not having an answer. What exactly was wrong with my pipetting technique? Did I pipette too quickly, or did I not press the plunger correctly?
The whole cell counter incident frustrates me because pipetting is something I have done in my past UROPs, yet I somehow haven’t mastered this straightforward skill. I feel bad for relying so much on my supervisor’s help. Having him help me is great, but depending on him is not sustainable as he has other work to do. I have to learn how to become independent in the lab setting. I feel overwhelmed when I consider the infinite number of possible mistakes I can introduce into my experiments because of my mediocre pipetting technique.
What’s more irritating is that I made a silly mistake almost at the end of setting up my 96-well plate of cells for a cell proliferation assay study. Instead of seeding my calculated volume of cell suspension into a new tube containing media, I pipetted media into my tube containing cell suspension solution. As a result, this cell solution volume is somewhat greater than my calculated value.
While the mistake doesn’t necessarily wipe out everything I did beforehand, I am worried that the cell concentration in each well may be higher than desired. There are also other worries I have now because of this error I caused, like whether each well has an even distribution of cells because I am not confident that the cell solution was mixed very well. But this is not a blog about biology experiment techniques, so I will stop there regarding the technical information.
Maybe I am exaggerating how bad the situation is, but I am so upset that I don’t know how to feel better. It’s only afternoon, but I have no idea how to make the rest of the day better. There is a MISTI Taiwan meetup today at 6:30 p.m., but I don’t feel like going anymore when all I can think about is how rough today’s lab work went for me. I want to return to my apartment right after work and stay inside my room. I simply want this day to end quickly so I can forget about it after falling asleep and then wake up with a fresh start.
I search some self-help articles online on what to do when one is in a bad mood, but I don’t like any of their advice. The articles suggested things like doing breathing exercises, but that doesn’t sound helpful. Other suggestions included venting it out, but I am not close with anyone in the lab. Playing sad music doesn’t sound that appealing either, but writing a blog post about this sounds like a good idea.
For some time, I close my eyes and put my head on the table to rest. I planned on taking a nap, but I couldn’t with this problem inside of me that kept expanding like a balloon. It’s only the first week of July. I don’t want lab errors like these to keep happening for the next six weeks. My head hurts when I consider this potential reality. My brain hurts even more when I think about having to plan and perform the western blot experiment in addition to this assay later this month.
After my short rest, I think about the articles I read. While they weren’t that helpful, one nice thing is that they made me think of how I would talk to a friend experiencing the same problem. To be fair, I have only been in the lab for a month. While four weeks is a decent amount of time, mastering lab techniques probably needs more time. I have done UROPs in the past, but this one is a lot more demanding.
As much as I wish things went smoothly, lab work doesn’t work like that. There are so many failures in the beginning, especially when one isn’t super familiar with the protocol. There are so many possible factors responsible for causing a lab error that it’s hard to pinpoint the actual reason. Other times, it’s carelessness or lack of attentiveness that causes lab mistakes, which is probably more frustrating since those are preventable. If something worked the first time, I would have to say that you are lucky, or you are extremely good at designing the experiment to the point of eliminating all possible sources of error.
Lab mistakes are not ideal, but mistakes happen to everyone when doing lab work and they are an essential part of the learning process. Also, humans are not robots, so it’s unreasonable to expect perfection. I just need to accept that as a fact of life if I don’t want to be that frustrated again.
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