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Use an ultra-clean workbench during cell culture operations
日期:2024-12-04 01:12
浏览次数:358
摘要:Cell culture is also called cell cloning technology, which refers to a method that simulates the internal environment (sterile, suitable temperature, pH, and certain nutritional conditions) in vitro to make it survive, grow, reproduce and maintain its main structure and function. There are mainly animal cell culture, plant cell culture, and microbial cell culture. Cell culture technology is an important and commonly used technology in cell biology research methods. Through cell culture, a large
Cell culture is also called cell cloning technology, which refers to a method that simulates the internal environment (sterile, suitable temperature, pH, and certain nutritional conditions) in vitro to make it survive, grow, reproduce and maintain its main structure and function. There are mainly animal cell culture, plant cell culture, and microbial cell culture. Cell culture technology is an important and commonly used technology in cell biology research methods. Through cell culture, a large number of cells can be obtained, and cell signal transduction and cell signal transduction can be studied. Anabolism, cell growth and proliferation.
Bacterial culture
Objective: Establish a set of applicable culture procedures to expand the culture of cells in vitro under pollution-free conditions.
Steps:
1. Ultra-clean workbench is irradiated by UV lamp for 30 minutes (According to the actual situation, the irradiation time can be appropriately extended or shortened, but a longer time is always safer than a shorter time.)
2. Incubate the culture medium, PBS, and pancreatin in a 37°C water bath (how much to use each time, and how much to incubate, which can save incubation time and extend the life of the drug.)
3. After 30 minutes of irradiation on the ultra-clean workbench, turn off the UV, turn on the lighting, turn on the exhaust air for 10 minutes, and then enter the cell culture room again. (Note: You must turn on the exhaust for 10 minutes before performing the operation, so as to ensure that the circulating air is sterile. At the same time, it will also prevent the bacteria from blowing directly into the human respiratory tract, which also has a certain protective effect on the human body)
4, wear kouzhao and gloves (conditionally ** wear a hat, but it does not matter, our laboratory did not wear it)
5. Wipe the test items with 70-75% alcohol, and then put them into the ultra-clean workbench.
6. Pay attention to the following points during operation: try to operate near the flame of the alcohol lamp, but if there are cells in the tube, there must be a certain distance from the flame, otherwise the cells will be burnt to death; do not reuse the pipette (that is, after sucking it once) , Just replace it with a new tube); do not operate over an open container; do not rub too much alcohol on your hands, otherwise you will easily burn your hands when you are close to the alcohol lamp (I think many people have been burned);
Clean bench
The operation is actually very simple, but you must be careful. Especially in step 2, many people don’t pay attention. Cells are usually cultured at 37°C. If the temperature of the added PBS or trypsin is too low, the cells will be injured. Although this damage is not noticeable in a short time, it will take time. It will show up when it grows.
Bacterial culture
Objective: Establish a set of applicable culture procedures to expand the culture of cells in vitro under pollution-free conditions.
Steps:
1. Ultra-clean workbench is irradiated by UV lamp for 30 minutes (According to the actual situation, the irradiation time can be appropriately extended or shortened, but a longer time is always safer than a shorter time.)
2. Incubate the culture medium, PBS, and pancreatin in a 37°C water bath (how much to use each time, and how much to incubate, which can save incubation time and extend the life of the drug.)
3. After 30 minutes of irradiation on the ultra-clean workbench, turn off the UV, turn on the lighting, turn on the exhaust air for 10 minutes, and then enter the cell culture room again. (Note: You must turn on the exhaust for 10 minutes before performing the operation, so as to ensure that the circulating air is sterile. At the same time, it will also prevent the bacteria from blowing directly into the human respiratory tract, which also has a certain protective effect on the human body)
4, wear kouzhao and gloves (conditionally ** wear a hat, but it does not matter, our laboratory did not wear it)
5. Wipe the test items with 70-75% alcohol, and then put them into the ultra-clean workbench.
6. Pay attention to the following points during operation: try to operate near the flame of the alcohol lamp, but if there are cells in the tube, there must be a certain distance from the flame, otherwise the cells will be burnt to death; do not reuse the pipette (that is, after sucking it once) , Just replace it with a new tube); do not operate over an open container; do not rub too much alcohol on your hands, otherwise you will easily burn your hands when you are close to the alcohol lamp (I think many people have been burned);
Clean bench
The operation is actually very simple, but you must be careful. Especially in step 2, many people don’t pay attention. Cells are usually cultured at 37°C. If the temperature of the added PBS or trypsin is too low, the cells will be injured. Although this damage is not noticeable in a short time, it will take time. It will show up when it grows.