Surgery Godfather

Chapter 2231: Chapter 1455: An Unexpected Place

Yang Ping’s gaze lingered on the microscope’s eyepiece for a long time.

The cell state was indeed not bad, with good adherence, normal refraction, no obvious signs of apoptosis or aging. But he always felt something was off, a sense of discord that couldn’t be put into words, like a dish with all ingredients added, yet missing something intangible in taste.

He straightened up and turned to Weber: "Let me see your experiment logbook."

Weber handed over a thick hardcover book, with a label on the cover bearing his name and experiment number. Yang Ping opened it, starting from last week’s record of the "beautiful data."

He read slowly, carefully checking each page. Batch numbers of reagents, cell generation, culture conditions, treatment times, detection methods... each entry was meticulously recorded, with handwriting as neat as print.

"The stem cells you used last week, which generation were they?" Yang Ping asked.

"Fourth generation."

"And these three repeats?"

"Also fourth generation," Weber said, "same batch of cells, simultaneously revived, passaged, and treated."

Yang Ping flipped back to the first page of the record book, his gaze landing on the batch information of the original cells.

"What about the original cells?" he asked, "Were the ones used last week the same batch as these three repetitions?"

"Same batch," Weber said, "all prepared in March. Original cells, stem cells, serum, all batches are exactly the same."

Same batch of serum, same batch of original cells, same batch of stem cells, same operator, same experimental plan. Three repetitions, yet only one succeeded. This can’t be an operational error; with Weber’s level of skill, he wouldn’t make such a basic mistake. It’s also not due to batch differences, as all batches are completely identical.

Then there is only one possibility: the problem isn’t at the operational level, nor at the batch difference level, but in an even more unexpected place.

"So," Weber slowly said, "last week’s data is real, and these three failures are also real. The same conditions led to different results. This means..."

"It means you encountered the most challenging type of problem in research," Yang Ping picked up his words, "where all visible and tangible variables are controlled, yet the results are still irreproducible. This suggests there is a variable in your experimental system that you haven’t discovered or even thought of. It’s not a batch issue, nor an operational issue, but something deeper."

This deduction left the three of them silent.

Tang Shun furrowed his brows in thought: "What could it be? Temperature? Humidity? Carbon dioxide concentration in the incubator? We monitor all these."

"Monitoring doesn’t equate to true control," Yang Ping said, "you monitor the set values of the equipment, not the actual environment the cells perceive. The temperature probe in the incubator is placed in the water bath, showing the water temperature, not the actual temperature inside the culture dish. Every time the door is opened to handle cells, the temperature fluctuates, humidity changes, and carbon dioxide concentration drops. These small fluctuations are inconsequential in most experiments but could be the key to success or failure in some sensitive systems."

Weber’s eyes narrowed slightly, he understood Yang Ping’s point.

"Are you saying that during last week’s experiment, the state of the incubator was different from these times? For example, last week no one was frequently opening the door, but this time someone was using the incubator next door, causing frequent temperature fluctuations?"

Yang Ping said, "It’s possible, but it might not be an incubator problem at all. It could be the pH of the culture medium; if it’s stored too long in the four-degree fridge, it becomes alkaline, and subtle pH changes can alter cell behavior. It could be cell confluence; errors in cell counting during passage could lead to varying cell densities when seeded. It might even be the treatment time; although the same operator, last week’s treatment was done in the morning, and these were done in the afternoon, with cells staying in the incubator a few extra hours, altering their state."

Yang Ping paused, looking at Weber: "In fundamental research, we pursue certainty, where the same conditions yield the same results. But biological systems are never certain. Cells are alive, and living things fluctuate, have noise, and unpredictability. The experiment you do today and the one you do tomorrow can never truly be the same. What we can do is not eliminate this uncertainty, because we simply can’t, but to understand it, quantify it, and control it."

As a top-tier scientist, Weber was well aware of this principle. He took a deep breath and then slowly exhaled, "I’ll start from scratch, breaking down every aspect of this experiment. For every potential problem, I’ll set controls. I’ll put a temperature recorder beside the culture dish to monitor the temperature fluctuations in real time. I’ll measure the pH of each batch of culture medium, not just rely on batch numbers. I’ll not only count cells but also photograph and record their confluence. I’ll time the treatments down to the minute. I have to find that hidden variable."

"Exactly!" Yang Ping said, "Don’t assume conditions are the same; prove they are the same."

Weber suddenly remembered something. He said, "Actually, I’ve encountered a similar situation, about ten years ago, while researching neural stem cell differentiation. There was a crucial experiment, the first time I did it, the results were excellent, but for three whole months afterward, I couldn’t reproduce it. I controlled all the conditions, cells, reagents, incubators, procedures, exactly the same. Yet the results just wouldn’t come; I almost went crazy at the time."

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