Perspective: Patching My Patch-Clamping
The first time I worked in a neuroscience lab was in 2014 when I joined a fundamental research group in Amsterdam and got trained in the “patch-clamp” method.
It was my first time doing wet lab work, so I had various opportunities to mess up. I fully took advantage of each one of them.
The patch-clamp technique is used to study the electrical properties of neurons and works as follows: A glass tube much smaller than the eye of a needle in diameter, known as a micropipette, is used to remove a tiny “patch” of the cell membrane in order to access intracellular space. The micropipette is filled with an electrolyte solution and there is a thin electrode that records the electrical current in the nerve cell. This electrode is connected to an amplifier that amplifies the signal, which, in turn, is connected to a digitizer that digitises the signal.
If all this sounds too complicated to understand, it certainly is: it took me about six months to start having a remote idea of what I was doing. Nevertheless, I was very excited. An entire new microworld was unfolding in front of my surprised eyes.
People’s voices were filled with fear and respect when they talked about the micropipette machine. They warned new students, multiple times a day, that they should not, under any circumstances and by no means, ever, until the end of times, touch the rest of the settings except the temperature.
On my first day at the lab, Victor, another student who was one year ahead of me, taught me how to make my micropipettes. He led me to the room where the micropipette-making machine was and explained that the only thing that I had to do was put a piece of glass inside the machine, program it to the right temperature, and then the machine would heat up, melt the glass and then pull it apart from the two sides, creating micropipettes with invisibly thin tips.
This sounded straightforward, but there was something sacred about it. People’s voices were filled with fear and respect when they talked about the micropipette machine. They warned new students, multiple times a day, that they should not, under any circumstances and by no means, ever, until the end of times, touch the rest of the settings except the temperature. This would cause a great lab catastrophe and it might even break the machine, which was very expensive to fix. This terrified me, so I listened obediently and never deviated from the exact instructions that I was given: pick up a glass, put it inside, set up the temperature, and let the machine do the rest. This way nothing bad will ever happen.
Victor had his set-up right across mine, as both of us were based at the corner of the patch-clamp lab, and we shared a long desk. He was tall, skinny, and perhaps slightly awkward in his movements. His eyes were wide and blue and if you looked carefully into them, you could find hints of kindness and sadness, well-hidden behind the serious look. In the beginning, we thought that we would not get along. I thought that he was a bit too shy, maybe even arrogant. He thought that I was a bit intrusive, and certainly too loud. We would definitely not get along.
The next step, when you already had your micropipettes, was getting used to moving them towards the cells using joystick-like micromanipulators that allowed the execution of very fine and delicate movements, which are essential for the success of the operation. Neurons are apparently very easy to kill – I learned this the hard way.
Moving the micropipette towards the cells in a delicate manner is a tough task. Most students spend the first couple of days or even weeks breaking one pipette after the other, hitting them with force on the cell plates, killing several cells each time, until they finally get it. I was not an exception. I broke all my pipettes into pieces with admirable speed, then went to the machine to make some more and quickly came back to break those as well. Every broken pipette was a blow to my self-esteem. Every dead cell planted a doubt regarding whether this internship had really been a good idea, whether this master’s program had really been a good idea, whether every single life decision so far had really been a good idea. Victor tried to comfort me, saying that it happens to everyone, and I should not take it personally. This was already difficult for my impatient nature and detecting a tone of amusement in his comforting words was not very helpful.
Every broken pipette was a blow to my self-esteem. Every dead cell planted a doubt regarding whether this internship had really been a good idea, whether this master’s program had really been a good idea, whether every single life decision so far had really been a good idea.
Eventually, I got used to it and started patching with relative success. Life started feeling nice in the microworld Victor and I had created in our little patch-clamp corner. We spent hours every day back-to-back, each one of us patching our cells while talking about science, free will, the past, and the future. We divided the tasks so that one could make the pipettes for both of us, while the other would go to the chemistry lab and prepare the solutions. However, most of the time we would spend double the time and do everything together because we didn’t want to interrupt an interesting conversation- or most probably an endless argument about how things in the world are or how they should be. We listened to Arctic Monkeys and Radiohead, and when a patched cell looked particularly healthy, we would ask each other to look at it with pride and joy. Sometimes we even gave the cells names, like Balthazar or Archibald, and laughed until someone else from the lab would tell us to keep it down.
One day, patching just stopped working. Suddenly, we both became incapable of getting it right and cells started dying one after the other, fogging both the transparent solution and our moods. When we got the cells out of the incubator, they looked happy and healthy, but by the time we tried to patch the first one of the day, everything was dying. We tried again and again and again, doing everything from scratch with extra care, we even started putting on the lab coats we never normally wore. Nothing could make the cells happy again and days went by without us being able to figure out why. These were not good days for our self-regard. What could we have possibly done to make the gods of patch-clamp so angry?
Working hours became silent. We thought of talking to our supervisors and decided not to get them involved. We had to solve this ourselves. I was grateful that they had accepted me in the lab without any prior experience and my supervisor was nothing but kind to me. I did not want to disappoint her by admitting that I am getting her cells killed day after day, and I don’t have the slightest idea of why this might be. I started noticing that the light in the lab was too white, the cold outside was getting very thick, and the constant rain and fog were washing out the colors of the surroundings.
One of these gloomy Amsterdam days, I was sitting on the lab bench with the bottle of artificial cerebrospinal fluid (ACSF) open, ready to pour some drops on my cell plate before placing it under the microscope. Victor was by my side, silently waiting for his turn. A momentary distraction made me pour some of the solution on the table when an undergraduate student walked in and grimaced: “What is this smell in here? It smells like bleach!”
What could we have possibly done to make the gods of patch-clamp so angry?
Victor and I looked at each other and immediately knew. Somehow, we had mixed up the labelling of the bottles and labelled the bleach we used for cleaning the set-up at the end of each day as ACSF, and the ACSF we bathed our cells into for keeping them healthy, as bleach. We went into an unstoppable, hysterical laugh that brought tears to our eyes. The mystery was solved.
This was not my biggest “goof” in science. I messed up several times after that, during the same internship and during the years after – I even messed up with the micropipette god-like machine: I made the hubris of changing its settings. Overall, I messed up in all kinds of original or even innovative ways. However, this first failure happened at a time when the absolute lack of experience made it feel catastrophic. Every mistake after that made me feel progressively less desperate, only a tiny bit less each time, until I finally understood that messing up is part of the learning process. Isn’t that what science is, falling forward?
With Victor, as it might be obvious by now, we did get along. We shared more failures, but also successes, during the months we spent in our patch-clamp corner and after that. He’s always been present in my life ever since. That is also what science is: falling forward, together.
