21: Chapter 21 The First Step to Implementation (Part 2)

After one practice session, Chen Youhua leaned back in his chair to relieve the fatigue in his eyes and shoulders.

"Your operational proficiency has improved,"

Wang Zhe commented.

"But you're too tense.

The equipment has a certain degree of fault tolerance; the key is to understand the purpose and impact of each operation.

After the chips arrive next week, continue practicing on regular petri dishes first.

The goal is to be able to complete the entire imaging process stably and reliably, reducing human error."

Chen Youhua's figure—handling failed samples, tuning parameters, and checking records in a corner of the lab—became an increasingly common sight.

He was a man of few words, putting most of his energy into solving the problems at hand.

Initially, some senior students would occasionally ask about his progress out of curiosity, but now they were used to it.

Occasionally, someone passing by would glance at the suboptimal images on his screen and ask a simple question.

"Chen Youhua, how's the data today?"

Chen Youhua would usually reply, "Still optimizing the conditions."

Once, Senior Sister Li saw petri dishes marked as failures and printed images lying by Chen Youhua's desk.

She picked up one of the images.

"Is the experimental record for this failed data complete? Serum batch number? Specific staining duration? Laser power and scanning speed during imaging?"

"It's recorded in the notebook."

Chen Youhua pointed to a lab notebook nearby, filled with dates, steps, and parameters.

"Good."

Senior Sister Li nodded.

"Failure isn't scary; the key is to find patterns and reasons from the records.

Regularly reviewing and analyzing this information is more effective than blindly repeating experiments."

It was nearly eleven o'clock when he returned to the dormitory that night.

Zhao Lei, wearing headphones and playing games, caught a scent.

"Chen Youhua, that disinfectant smell on you is pretty strong."

Wang Pengfei stuck his head out. "Want to play? It's a new season."

Chen Youhua replied while changing his clothes, "No, I need to look at data."

After washing up, he sat at his desk, turned on his computer, and pulled up the daytime experimental parameter records for comparison.

Li Fan looked up for a moment, said nothing, and continued reading.

The dormitory was filled with game sound effects and the clicking of keyboards.

After returning to the lab the next day.

Tuning the cell status was like fumbling through a fog.

Poor cell adhesion, low fluorescent labeling efficiency, environmental fluctuations affecting cell activity...

Facing the detailed failed data in his lab notebook, Chen Youhua troubleshot the variables one by one.

Increasing serum concentration? Limited effect.

Lowering seeding density? Cell dispersion worsened the adhesion problem.

Trying different batches of Fluorescent dyes? Background interference increased significantly in some batches.

"Is the culture medium preheated sufficiently? Is the actual temperature of the water bath constant?"

Wang Zhe reminded him as he passed by.

"The digestion time during cell subculturing also needs precise control; over-digestion damages the cells."

Chen Youhua checked the records and found that the digestion time had indeed exceeded the standard once.

This might have been one of the reasons for the poor state of that batch of cells.

In subsequent operations, he strictly timed everything.

After multiple parameter adjustments.

He optimized the serum ratio, precisely controlled cell density and digestion time, screened dye batches, and determined the optimal staining and recovery times.

In one experiment, Chen Youhua observed that most cells adhered well in the petri dish, showing normal morphology, and the labeled microtubules within the cells displayed clear, stable fluorescent signals.

"The state is good this time." Wang Zhe noticed it too.

Chen Youhua cautiously set long-term imaging parameters and started the scan.

The first half-hour went smoothly.

However, about an hour later, he noticed blurring at the edges of the field of view.

Inspection revealed that condensation had formed on the inner side of the objective lens due to local temperature changes.

He paused the sequence, carefully cleaned the objective lens, and refocused and repositioned.

This process was time-consuming and could affect the cell status.

More troublesome was that after restarting, some microtubules originally in the center of the field had moved out of the observation range due to the cells' own activity.

The precision of environmental control became a challenge once again.

Through repeated setbacks and adjustments, Chen Youhua gradually accumulated experience.

He developed the habit of checking for environmental interference sources in advance, recording the actual temperature of the water bath, placing temperature probes near the cells for real-time monitoring, and confirming the CO2 supply before imaging.

This afternoon, he seeded the cells again, optimized the staining conditions, and after confirming environmental stability, started a new round of 3-hour imaging.

He forced himself to quickly check the imaging status periodically.

After the sequence was completed, Chen Youhua opened the saved image files.

Dragging the timeline, the cells on the screen remained stable, and the internal microtubule fluorescent signals were clearly visible most of the time.

Despite slight field-of-view drift and local background interference that made tracking difficult in some areas, he had obtained his first segment of relatively complete live-cell microtubule dynamic imaging data suitable for preliminary analysis.

There were no obvious emotional fluctuations.

Chen Youhua silently backed up the data and then tried to perform automatic tracking analysis using TrackMate software.

The results were still unsatisfactory; the software made many identification errors, and the trajectories were severely broken.

But at this moment, looking at the messy tracking lines overlaid on the raw images, his state of mind was much calmer than before.

At least he had usable raw data; software analysis was the next hurdle to overcome.

The pressure from continuous high-intensity work, intense mental focus, and repeated troubleshooting began to show.

While operating in front of the microscope once, Chen Youhua's vision suddenly blurred briefly, and his hand control became unsteady, nearly knocking over the nearby Fluorescent dye.

He quickly steadied the bottle, his heart racing.

"What's wrong?" Wang Zhe noticed something was off and walked over.

"My eyes are a bit blurry." Chen Youhua pressed his temples, his voice revealing exhaustion.

Wang Zhe scrutinized the dark circles under his eyes and his slightly pale complexion.

"What time did you go to bed last night? Two? Or even later?"

Chen Youhua did not deny it.

"Reckless!"

Wang Zhe's tone was serious.

"Scientific research is a marathon, not a sprint.

With your current state, you can't concentrate, and your operational precision has dropped. Not only will you fail the experiments, but you might also damage the equipment or even cause safety issues! Can you take responsibility for that?"

Chen Youhua remained silent, the feeling of physical exhaustion heavy and real.

"Now, go back and rest immediately!"

Wang Zhe pointed to the door.

"Pack your things and go back to the dorm to sleep! You are not allowed to touch any experiments tonight! Put your phone on silent and deal with anything, no matter how big, tomorrow!"

Chen Youhua looked at the unfinished practice, felt his body's strong protest, and finally nodded.

He saved the data, turned off the instruments, and took off his lab coat.

Walking out of the lab building, the night breeze brought a hint of cool, but the fatigue ran deeper.

Back at the dorm, Zhao Lei and Wang Pengfei were gaming. Zhao Lei took off his headphones.

"So early today? That's a first!"

Chen Youhua gave a vague response and headed straight for his bed. Even washing up felt like an effort.

He collapsed onto the bed, the sounds of his roommates' gaming seeming to be behind a barrier.

Before sinking into sleep, the lingering thought was of that bottle of expensive green Fluorescent dye that had nearly tipped over.

The first step of scientific research practice, accompanied by setbacks, tuning, and physical exhaustion, had truly and concretely begun.