Thursday, February 11, 2021

(The second day of antibiotic treatment)

8:10 AM — I begin my day by inspecting the setup, recording measurements from the scales, and noting visual observations of the reactor, such as the overall appearance of the culture. Today, while the setup seems stable, the culture appears rather lackluster.

Upon checking the fluid density in my reactor, I notice minimal growth overnight, which is not unexpected given that I initiated antibiotic treatment yesterday—designed specifically to eliminate bacteria.

8:20 AM — I prepare to take samples. This involves retrieving items from the fridge, gathering pipettes, and sanitizing my workspace with 96% ethanol—just the usual routine.

8:30 AM — With everything ready, I start the sampling process. To avoid contamination, I work under a flame whenever I extract fluid, which helps to sterilize the surrounding air and keep outside microbes at bay.

The following thirty minutes involve transporting a bucket of ice containing my samples from room to room, utilizing various machines. However, while my samples are centrifuging, I notice a drop in pressure in one of my reactor tubes. Panic mode activated.

The medium is pumped into the reactor using a hydraulic pump, and with the pressure lost, no medium is entering the system. This presents several issues, but I won’t delve into the details.

Fortunately, I managed to resolve the issue in about ten minutes without fully understanding what went wrong or how I fixed it. Quite frustrating.

9:05 AM — I’m feeling the pressure. Due to the earlier issue, I was unable to prepare the antibiotic treatment, and my samples need to be stored at -20°C/-80°C immediately after centrifugation.

9:15 AM — After some frantic rushing, I successfully store my samples and get ready to introduce antibiotics into the reactor, which has to be done at a precise time. I have reservations about spiking the reactor with antibiotics.

After ten days of nurturing a stable community, I am now at risk of destroying it. I worry about potentially annihilating all the bacteria, which contradicts my experimental goals. If that happens, I'd need to start over and revise my experimental design. I want to avoid that.

9:20 AM — No time to rest yet; I must finish sampling. I collect samples for various analyses, some of which need immediate freezing, while others can wait a bit longer.

As I continue to move around, weighing and measuring, I make sure to avoid burns from the 105°C and 550°C ovens. Once I finalize today’s samples, I also prepare those from the previous day for the next steps.

10:00 AM — For the first time today, I finally sit down since leaving home at 7:45. I begin entering the morning's data into my Excel sheets only to discover that my reactor received less medium overnight than expected—suggesting tubing issues persisted longer than anticipated.

There’s nothing to be done now; data is rarely perfect. As long as I document all observations, I can at least clarify any anomalies when reviewing results. As a perfectionist, this stresses me out immensely.

I strive for clean data without external interruptions. Yet, in a setting where machines and humans are involved, errors—both mechanical and human—are inevitable.

The antibiotic treatment is visibly impacting my bacterial population, as evidenced by my observations and real-time gas emissions data. I take some time to ponder the situation and review relevant research literature.

11:00 AM — I return to the lab to inspect the tubing, feeling a bit paranoid that something might be amiss again. Thankfully, everything appears to be functioning properly.

11:15 AM — I encounter one of the lab technicians, and we discuss the tubing issue. If the pressure was lost, it likely resulted from a "leak," allowing air to enter instead of my medium.

It turns out that one of my tubing connectors was not airtight when adjusted in a particular manner. A piece of tape with a "DO NOT TOUCH/MOVE!!!" note should fix the problem… and it did.

12:00 PM — Lunchtime, just in the nick of time! I was beginning to feel hungry. Eating or drinking in the lab isn’t allowed, and I often forget to snack while working in the office. Thankfully, I’ve got hydration under control these days. I find myself staring at my colleagues, feeling fatigued.

1:00 PM — Back to the lab, where I booked time in the "kitchen" to prepare medium. This involves weighing various chemicals and filling 10/20L bottles, which I also have to lift on and off a cart. Arm workout of the day: check!

2:15 PM — In the office, I troubleshoot an issue from the previous day that didn’t yield the desired results, though I’m still uncertain about what went wrong.

2:20 PM — I decide to approach it differently this time, heading back to the lab for a redo. Let me tell you, nothing ever works on the first attempt in the lab. Never.

3:15 PM — Another round of sampling, much like this morning. It takes about an hour to complete all the various samples. Afterwards, I prepare tubes and labels for the next day. I often underestimate how much time this takes.

4:00 PM — I’m feeling exhausted, my back is aching, and my focus is waning. Danger zone. I still have tasks to complete in the lab before I can leave.

4:30 PM — I take a necessary tea break to rehydrate and recharge a bit. I end up reviewing my data, so it’s not quite a break. Technically, I’m still working—just a break from the lab.

5:00 PM — Back in the lab, I quickly prepare for the second antibiotic spike at 5:15 PM. I conduct a final check to ensure everything is functioning as intended, sanitize my workspace with 96% ethanol, and bid goodnight to my cultures.

As I exit the building at 5:30 PM, the fresh air feels rejuvenating. The bioreactor room can be quite odorous, and it feels rewarding to head home after a long day of hard work.

Often, I’m too fatigued to log my afternoon entries into my Excel sheets, so I tackle that once I’m home and have had a snack. Additionally, I check my online data from the reactor every couple of hours to monitor gas profiles and ensure nothing is amiss—though I wouldn’t necessarily see an explosion on the profiles, nor could I do anything about it if it occurred.

Tomorrow will likely mirror today, yet it will present its own unique challenges. Since cells don’t take weekends off, I’ll need to return for an hour of sampling—nothing too arduous. Lab work with living cultures demands significant commitment.

A Journey of Continuous Adaptation

From my experience, lab work can be physically taxing; one can easily surpass 10,000 steps on a busy day. Then, there’s the mental aspect to consider.

No two lab days are alike, which is exciting yet requires constant problem-solving and adjustments. Tasks often take at least three times longer than planned. My supervisor often refers to Murphy’s Law: “Anything that can go wrong will go wrong.”

Nonetheless, the thrill of research is captivating. The rush of achieving your goals or discovering something unexpected is exhilarating, akin to finding the final piece of a puzzle.

This encapsulates what it means to be “in the lab” for me: darting around to resolve issues, performing mundane tasks like labeling tubes, stressing over uncontrollable variables (if you’re like me), experiencing pure joy when things align, and the excitement of uncovering new knowledge—the delight of beginning to grasp complex concepts.

It’s reminiscent of being a curious child in a wondrous playground, albeit one filled with various compounds and machines that could be hazardous or even explosive. I truly never know when I’ll make it home.

If you’ve read this far, I’m genuinely impressed! Feel free to let me know if you found this interesting and if you’d like to learn more about my research. I’d be delighted to share more details!