"Your cells were contaminated. I got rid of them. Try again." This was the series of hope-depriving statements repeated to me many times during my seven month lab internship. I was working at a neuroscience lab at Vanderbilt University. I was attempting to culture neurons on graphene coverslips as a first step to showing that graphene based scanning of neurons would yield scans of a much higher resolution than traditional scanning methods. It sounded easy enough when I was first assigned the project, but, as I would soon find out, experimental science never goes right the first time, or even the first ten times.
My first challenge was to quickly learn how to isolate the hippocampus, an extremely slim piece of tissue, from a baby mouse's brain smaller than a dime. This involved a series of delicate surgical procedures that normally involves months, if not years, of training. Despite the facts that I did not receive this training and that I was a high school student, my lab members quickly expected me to be able to isolate the hippocampus on my own and perform the cell culture. Despite my pretensions, this was to be the easiest aspect of my time at the lab.
Given that isolated neurons lack a functioning immune system, even the slightest bit of bacterial contamination would result in complete neuronal death. For this reason, the most stringent sterilization procedures were put into effect. Having never been forced to work in a confined space, I often made careless errors in keeping all of my materials sterile. After each overnight incubation, I would rush to the storage chamber and examine my cells, only to find that symbolic cloudy mist that meant that I had yet again failed.
My PI refused to let me give up, and occasionally monitored me to ensure that I was adhering to all protocols. Eventually I became accustomed to the stringent procedures and I knew that I wouldn't fail the next time. I harvested the neurons, treated them, and plated them onto coverslips, just as my PI had taught me only days previously. I treated every action as a possible potential for failure, and I made sure that I never slipped up. Upon completion, I nervously stored the cells into the storage chamber overnight. The next day, I sprinted to the storage chamber. Success! The portentous mist was absent, and I had finally succeeded in a cell culture. I thought that the hard part of my experiment was behind me. I was very wrong.
My next agonizing ritual was to apply three small wax dots to a coverslip for a neuron support structure. Essentially, this entailed melting the wax on a hotplate and transferring drops to the coverslip using a pipette. Again, all of this had to be done within the flowhood to ensure sterility. Upon reading this, I thought to myself that this was utter child's play compared to the cell culture. Then I tried it. My first attempt at transfer resulted in the wax hardening in the pipette, rendering both unusable. The wax was obviously too cool, so I heated it further. My second attempt at transfer saw the wax drop spread throughout the entire coverslip upon impact, as the wax was too hot. It turned out that there was an absurdly small range of temperatures that would allow for successful wax deposition, and the hotplate that I was using divided temperatures into numbers from 1 to 6, not helping my problem any. I thus quickly realized that my endeavor would be based on pure trial and error. One hour, thirty pipettes, and fifteen coverslips squandered, I finally achieved my task. One coverslip was prepared. Only 23 more to go...
I learned many things from my tedious days in the lab. Foremost, science in the real world is nothing like science taught to you at school. You are not hand-fed the answers; you have to painstakingly work to find them yourself. An additional positive experience was the ability to work with professionals in one of my fields of interest. This offers an accelerated and more in-depth learning process that is at a much higher level than traditional high school doctrine. But the most important thing that I learned from my time at the lab is brought to you from the wise mind of William Edward Hickson: "If at first you don't succeed, try try again."
My first challenge was to quickly learn how to isolate the hippocampus, an extremely slim piece of tissue, from a baby mouse's brain smaller than a dime. This involved a series of delicate surgical procedures that normally involves months, if not years, of training. Despite the facts that I did not receive this training and that I was a high school student, my lab members quickly expected me to be able to isolate the hippocampus on my own and perform the cell culture. Despite my pretensions, this was to be the easiest aspect of my time at the lab.
Given that isolated neurons lack a functioning immune system, even the slightest bit of bacterial contamination would result in complete neuronal death. For this reason, the most stringent sterilization procedures were put into effect. Having never been forced to work in a confined space, I often made careless errors in keeping all of my materials sterile. After each overnight incubation, I would rush to the storage chamber and examine my cells, only to find that symbolic cloudy mist that meant that I had yet again failed.
My PI refused to let me give up, and occasionally monitored me to ensure that I was adhering to all protocols. Eventually I became accustomed to the stringent procedures and I knew that I wouldn't fail the next time. I harvested the neurons, treated them, and plated them onto coverslips, just as my PI had taught me only days previously. I treated every action as a possible potential for failure, and I made sure that I never slipped up. Upon completion, I nervously stored the cells into the storage chamber overnight. The next day, I sprinted to the storage chamber. Success! The portentous mist was absent, and I had finally succeeded in a cell culture. I thought that the hard part of my experiment was behind me. I was very wrong.
My next agonizing ritual was to apply three small wax dots to a coverslip for a neuron support structure. Essentially, this entailed melting the wax on a hotplate and transferring drops to the coverslip using a pipette. Again, all of this had to be done within the flowhood to ensure sterility. Upon reading this, I thought to myself that this was utter child's play compared to the cell culture. Then I tried it. My first attempt at transfer resulted in the wax hardening in the pipette, rendering both unusable. The wax was obviously too cool, so I heated it further. My second attempt at transfer saw the wax drop spread throughout the entire coverslip upon impact, as the wax was too hot. It turned out that there was an absurdly small range of temperatures that would allow for successful wax deposition, and the hotplate that I was using divided temperatures into numbers from 1 to 6, not helping my problem any. I thus quickly realized that my endeavor would be based on pure trial and error. One hour, thirty pipettes, and fifteen coverslips squandered, I finally achieved my task. One coverslip was prepared. Only 23 more to go...
I learned many things from my tedious days in the lab. Foremost, science in the real world is nothing like science taught to you at school. You are not hand-fed the answers; you have to painstakingly work to find them yourself. An additional positive experience was the ability to work with professionals in one of my fields of interest. This offers an accelerated and more in-depth learning process that is at a much higher level than traditional high school doctrine. But the most important thing that I learned from my time at the lab is brought to you from the wise mind of William Edward Hickson: "If at first you don't succeed, try try again."
