This doctor has a system

Chapter 171. Chapter 169

The next day, when Qin Zhifeng was still busy in the ward, his teacher, Qiao Liangping, called him, "Come here now, let's report to Director Zhang."

Qiao Liangping thought about it all night and decided to give in. There was no other way. He was about to retire, but there were still so many disciples here. If Director Zhang really didn't teach martial arts, then they might be in a very difficult situation.

But don't underestimate the ability of the department director of a large tertiary hospital. As long as you are in this department, it is very easy for him to clean up. The promotion of the title alone can get you stuck.

"Old Qiao, what is this?" Director Zhang asked knowingly.

"Director Zhang, I'm confused." Qiao Liangping said regretfully, "I was jealous at the time, and that's why I talked nonsense in the association. I reflected on it in the past few days and felt that my problem was very serious. Come here to apologize to you."

"Look at you, what are you talking about." Director Zhang smiled and said, "I know you must also be thinking about the association, but it is indeed a little thoughtless, and this is not a big problem."

"Just correct it in time."

"Then I'll call Director Shen right now and admit my mistake." Qiao Liangping said, he was completely out of face, he had no choice but to stand at attention when he was beaten. Really soft.

"That's not necessary, I'll just tell her." Director Zhang kindly saved him some face.

"That's too much trouble for Director Zhang." Qiao Liangping lost his temper at this moment, "Zhifeng's leading the team..."

"This year is not enough, the quota is all given to Lao Yuan." Director Zhang said, "Wait until next year, I will sign for you first, and you will take it to the Medical and Education Department in a while."

After leaving the house, Qiao Liangping felt much better. "It's all the teacher's fault. It's okay to make enemies everywhere, and you've been delayed for another year."

"Just one year, I can afford to wait." Qin Zhifeng breathed a sigh of relief, but he didn't mean to blame the teacher in his heart.

Director Shen soon received a call from Director Zhang, "This old Zhang has a way." She smiled and said to Director Zhao, "That Director Qiao dared to rely on his old man in front of me, it's really embarrassing to him. "

"That's right, he had nothing to do with him, so he insisted on rushing up and beating him up. I've heard of Director Zhang's methods, and he must have been dealt with severely." Director Zhao said casually.
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On the side of Township X, Hu Jiaran was worrying in the laboratory, they encountered a problem.

The preparation of feeder cells for fusion is relatively smooth. The source of peritoneal macrophages is convenient and the preparation is simple. The most important thing is that the macrophages have the ability to remove dead cells and phagocytic debris. Therefore, peritoneal macrophages are generally selected for fusion. cells as feeder cells. .
"The 7 to 8-week-old female mice were sacrificed by dislodging their cervical vertebrae, soaked in 70% alcohol for disinfection, and then cut a small opening in the abdomen of the mouse with sterile surgical scissors in the ultra-clean workbench, and peeled off the surrounding skin with tweezers to expose the abdominal cavity;"

Discard the supernatant, add 1ml of complete medium to resuspend the cells, then dilute to the required volume, and add to the 96-well plate;

Even this step is complete.

The next step was to prepare the spleen cells of the immunized mice. Although this step was not easy, it did not cause too much trouble to Hu Jiaran and the others.

On the day of cell fusion, mice that had been immunized three days ago should be used as the source of immune spleen cells. Generally, the spleen volume of immunized mice is twice that of normal mice.

"Use high-pressure sterilized surgical scissors in the ultra-clean workbench to open a small opening in the lower left side of the mouse chest cavity, remove the spleen with forceps and place it in a dish containing 3-5ml of incomplete medium, and remove the surrounding connective tissue. Transfer the spleen to a wet 0.4-mesh sieve and gently grind it with a grinding rod, and add an appropriate amount of pre-cooled "Buwan."

It turned out that Hu Jiaran was very happy, but he didn't expect to encounter trouble during the integration.

The experimental procedure given by Gao Feng was to fuse myeloma cells and splenocytes of immune mice, but they conducted many experiments, all of which ended in failure.

"I think it's a matter of proportion." Yue Penghui said, "The proportion of the two cells plays a key role in the fusion."

"And the temperature!" said a man named Wang Wei. He has a postgraduate degree and has solid basic skills. Hu Jiaran has noticed this person for a long time.

"There is also the speed of stirring." Xu Bing, who has just been here for less than a month, also expressed his opinion, "Too fast stirring speed will cause spleen cells to rupture, and if it is too slow, it will easily coagulate. I think the speed is 1r/min. suitable."

Everyone talked in a hurry, the experiment got stuck at this point, it has been more than a week, and there is no progress at all, everyone is very anxious.

"What you said makes sense." Hu Jiaran concluded, "Then what should we do next?"

"How can I do it? Do it to death." Wang Liang shouted, "Go to bed at four o'clock, wake up at six o'clock, the King of Hades praised me for my good health!"

Hu Jiaran was suddenly very satisfied with this funny comparison, "Overtime pay is doubled."

After half a month.
"It's a two-day holiday, everyone, take a good rest." Hu Jiaran was very excited, but his voice clearly revealed weakness.

He braced himself and began to record the detailed procedure of the experiment:
Fully break up the precipitate, add 1ml of 32% PEG preheated at 50°C to the centrifuge tube at a constant speed (with a 1ml pipette, add within 5 minutes), and gently stir with the pipette tip during the process;
Continue to stir slowly with the gun head for 3~4 minutes;

Add 1ml of incomplete medium preheated at 37°C at a uniform speed (complete within 1 min), stirring gently with a pipette tip during this period;
Then add 3ml of medium preheated at 37°C at a uniform speed (complete within 3 minutes), and gently stir with a pipette tip during this period;
Add selective rehydration medium containing HAT within 24 hours after fusion, 50 μl/well;

Two days after rehydration, carry out the first half-change of medium, and suck out 60-80 μl of medium from each well, and the HAT content in the medium of the first half-change of medium is halved, 50 μl/well;

Two days later, perform the second half-change of medium, suck out 60-80 μl of medium from each well, and replace with HT-containing medium, 50 μl/well.

"You don't want to take a break and do it again." Qi Nannan persuaded from the side, "I'm really afraid that you will die suddenly."

"How is it possible?" Hu Jiaran didn't even raise his head, "I'm waiting. Also, Qi Nannan, can I give you a suggestion?"

"You said."

"That is, when you execute the mouse, can you not always show that perverted smile?" He said with disgust, "Even a pervert like Wang Liang says you are abnormal."

"You're sick, you guys!" Qi Nannan was unhappy, "I killed a few mice and who did I provoke? Then next time I'll keep my face blank."
-
Wang Liang and the others are making skewers on the street. There are a lot of people in the university town here, which is very lively.

"Look at these girls, they are really beautiful." Someone drooled, "There are many girls in the medical school, unlike our major, a department is full of old men, and it is impossible to find a match."

"Yeah, there are 6 people in my dormitory, all of them are single dogs after 4 years."

"Hey, I thought I could find a girlfriend soon after graduation, but who knew it would be more difficult than when I was in school, shit!"

"Didn't Mr. Hu say that after the project is completed, we can find a company for friendship?" Yue Penghui's eyes showed hope, "If you want me to say, nurses from the provincial hospital will do, and there must be many beautiful ones."

Speaking of girls, men will never lack topics in their mouths.

"It will be the 10th soon, and our wages will be paid soon." Someone mentioned this matter, "I roughly calculated, if it is true according to Mr. Hu, then I will get at least 3 this month."

"I'm about the same as you." Several people talked about this with great interest.

"It will be released immediately." Wang Liang revealed to everyone, "Yesterday I heard Mr. Hu called Manager Zhao and mentioned this matter, saying that it will be distributed as soon as possible to increase everyone's enthusiasm."

While speaking, he picked up his mobile phone and checked his bank account, "Damn it, it's been sent!"

Hearing what he said, everyone became uneasy, and started to check, "I remember that there is a function to remind the account? It means that someone sends money, and you can send a text message."

"That one has to go to the bank to activate SMS reminders, 2 yuan a month."

"Penghui, how much did you send?" Wang Liang asked.

"personal privacy."

"Your uncle's privacy, let me see it." Wang Liang reached out and took his mobile phone.

"43000?" Wang Liang was stunned, "No, why? You are 1 yuan more than me!"

"Penghui's key points have played a big role. How can I paddle every day like you." Someone laughed, "If I were the boss, I would have been kicked out by you."

Zhao Xingye made a special call to Gao Feng this morning, and the salary slip that Hu Jiaran reported really surprised him. Just these dozen or so people cost the company nearly 80 yuan a month, which is too exaggerated!

"How about I go over and check?" He suggested in a low voice.

"No need, they work a lot of overtime this month." Gao Feng said casually, "Staying up late is the easiest way to lose hair. These people will definitely go bald in the future."

"I see that you have been running back and forth recently. It's been very hard. If you can't do it, give yourself a raise." After finishing speaking, Gao Feng hung up the phone.

Zhao Xingye didn't recover for a long time, he fell into deep doubts, what happened to this boss?I always feel that the company is not run by him.

"Hurry up and start cloning the hybridoma cells." Gao Feng said to Hu Jiaran, "This step is very important. After the results come out, you should bring the data back to me as soon as possible. It will be used for writing articles."

Hu Jiaran originally wanted to slow down the progress appropriately. He was really afraid that these migrant scientific research workers would die suddenly one day, but since Gao Feng had spoken, everyone had to move quickly.

Whip the fast cow!

Cloning of hybridoma cells generally refers to cloning of antibody-positive wells.In actual experimental operations, there will be several or more clones, which may include the desired antibody-secreting cells, antibody-non-secreting cells, and other irrelevant antibody-secreting cells.The principle of cloning is to clone as early as possible the hybrid clones that are positive for antibodies.

Because the growth rate of non-antibody-secreting cells is faster than that of antibody-secreting cells, the growth of antibody-secreting cells will be inhibited, and eventually the competition between the two will result in the loss of antibody-secreting cells.

At present, the commonly used cell monoclonal methods mainly include the following two types: limiting dilution method and flow cytometry fluorescence sorting technology.

The limiting dilution method is currently the most widely used monoclonal selection method. It has obvious advantages, simple operation, relatively low demand for instruments and equipment, and is convenient for subsequent process development.

Its shortcomings are also obvious, such as long time-consuming, low efficiency, high cost and high error rate.Since this method is highly dependent on manual work, multiple rounds of limiting dilution are required to obtain a high monoclonal rate.

Flow cytometry fluorescence sorting technology selects the required single cells according to factors such as cell size, fluorescent dyes and cell surface characteristics. This technology has been widely used in single cell separation, rare cell sorting and other experimental processes.The advantage is that the operation is simple and the cycle is short, and the disadvantage is that the monoclonal rate is uncertain.

The monoclonal cells obtained according to the cell morphology will produce multiple cells with slight differences. Such differences will still cause excessive proliferation after passage and expansion, thus affecting the purity of the monoclonal.

Gao Feng provided Hu Jiaran with a new cloning method called high-throughput equipment automated screening. The advantages are less manual intervention and high throughput. The advantages are obvious, low cost and high efficiency. The requirements are relatively high.

Everyone in the laboratory had never heard of this method, and everyone looked at a loss.

"Brother Feng, I'm afraid this won't work. You've never heard of what you're talking about, and there's no such instrument on the market." Hu Jiaran called Gao Feng, "We've been researching for a few days, but we really don't have a clue."

"Don't panic, I'll go there now." Gao Feng was not in a hurry, and it was time for him to end himself at this time.

"First of all, we need to build an automated liquid processing system." Gao Feng said to a group of scientific research migrant workers, "This kind of system usually consists of multiple liquid processing units, which can automatically complete liquid pipetting, mixing, etc. according to the needs of different experiments. Operation. The automated liquid handling system has the characteristics of high precision, high speed, and high scalability, and can process multiple samples at the same time, which can rapidly improve screening efficiency and accuracy."

"There is no one on the market for now, but we can make one ourselves. It's not difficult. We just need to integrate some instruments together." He is full of confidence. He has done this step in the system space and fully achieved the goal. system requirements.

"This is the blueprint. You should contact Attorney Zhao to apply for a patent. Also, buy another 10 high-speed centrifuges." Before leaving, Gao Feng explained to Hu Jiaran, "I will make this within 2 weeks. There are programming and integration problems. Go straight to the professionals, don't be afraid to spend money, we can't waste any more time."

Hu Jiaran is under a lot of pressure. Judging from the current progress, three months is definitely not enough, and the results in four months will be very high-spirited.

Everyone was too busy, and the fortune tree bought when the work started was dead because no one watered it.

Looking at the dead tree with yellow leaves, Hu Jiaran sighed, "Isn't it said that the rich tree is drought-resistant? Why did it die so soon? This is not a good omen."

"Find two people to throw it away when you get off work."

"The fortune tree, if the tree dies, isn't there only fortune left?" Wang Liang said.

"It seems to be such a reason. Then put this first?" Hu Jiaran.

　Today is only one chapter, and tomorrow there will be [-] words.

　　
　
(End of this chapter)