First, the laboratory design 1. Cell culture is an aseptic technique that requires the working environment and conditions to be free from microbial contamination and other harmful factors. 1, ultra-clean workbench / biological safety cabinet Cell culture requires a highly sterilized environment. When performing cell experiments, it needs to be carried out in air at level 100. Then we need to purchase a clean bench or biosafety cabinet that can provide 100 levels of cleanliness. For most experiments, the ultra-clean workbench can fully meet the needs of the experimenter. As for the size of the clean bench, it depends on the size of the laboratory and the number of people to determine whether it is a single or a double, whether to buy one-sided or two-sided, and buy a few. If the operation of the experiment is biohazardous, you should choose a more advanced biosafety cabinet than the ultra clean bench. There is not much difference between the safety cabinet and the clean bench, but the safety cabinet protects the personnel and is suitable for experiments such as viruses or cell bacteria that are harmful to the human body. It should be noted that the placement of the clean bench or the safety cabinet should avoid the door, air conditioner and other places that disturb the nearby airflow. The buffer system used for cell culture is a sodium bicarbonate + HEPES system, so carbon dioxide is required to provide buffering capacity to maintain pH. At this point we need a carbon dioxide incubator to culture the cells. And because the carbon dioxide incubator usually does not have a refrigeration compressor, the user needs to determine that there is an all-weather air conditioning refrigeration in the cell room in the summer, otherwise the temperature inside the box will exceed the standard. In addition, the carbon dioxide incubator is also divided into a gas-sleeve type and a water-sleeve type. The so-called gas-sleeve type is a direct heating wire heating, the box body heats up quickly, but the temperature accuracy and the heat preservation effect are slightly worse; while the water jacket type uses the soaking The electric heating wire in the water heats the water jacket wrapped around the outer body of the box to achieve the effect of temperature control, and the temperature rise is slow but the effect of heat preservation is good, and even if the power is cut off for a short time, it is not afraid of affecting the cell growth in the box. Need help with an inverted microscope. For some fluorescent immune cell experiments (IF), an inverted fluorescence microscope is also required. Cell culture also requires ultrapure water system. Ordinary pure water and distilled water can not be used for cell culture, while ultrapure water is mainly used to configure some medium and reagent. The water of ultrapure water can be tap water or secondary pure. Water (good quality deionized water, resistance of about 10 megohms), in addition to ultra-pure cell culture water, the most important endotoxin has strict requirements, so when selecting ultra-pure water machine, it is necessary to select ultra-filter components. Model. Cell culture requires centrifugation to collect passage cells. The general rotation speed is between 1000 and 1500 rpm, so it is necessary to purchase a low-speed normal temperature centrifuge, and bring 10 ml, 50 ml and the rotor of the culture plate. A high-speed refrigerated centrifuge is also required to collect cell pellets or cell supernatants. To establish a cell bank, a liquid nitrogen tank is necessary. The frozen cells in a liquid nitrogen tank can generally ensure biological activity in the past ten years. As for the size of the liquid nitrogen tank, according to the size of the cell bank. And set. The cells need to be stored at -70 to -80 degrees overnight for cryopreservation and then placed in a liquid nitrogen tank, which requires a low temperature freezer. The cell house should be isolated from the outside world when it is constructed. This requires purchasing a transfer window to deliver the item, reducing the chance of contamination, and all the items entering the cell room are UV-sterilized in the delivery window to ensure the safety of the clean room. All kinds of cell laboratory utensils need to be sterilized before and after use. The sterilization conditions are generally 121 ° C for 20 to 30 minutes. This requires an autoclave placed in a clean room to ensure biosafety. In the cell experiment operation, various tips and pipettes are required, so it is necessary to configure the pipette. For cell experiments, oscillators and magnetic stirrers may also be needed to mix the liquid; some hard-to-clean stainless steel or glassware filled with cells require the help of an ultrasonic cleaner; if an immunological experiment is involved, ice is definitely needed. If the demand for bath and crushed ice is large, you need to buy an ice machine; in addition, the broken cells may require an ultrasonic cell disrupter; the last thing to mention is that if the cell house is large, you need to equip the cart, and move those when you experiment. Liquid media is always a tormenting thing. 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2, cell culture room and design principles are to prevent microbial contamination and harmful factors, requiring a clean working environment, fresh air, dry and smokeless dust.
3. The design principle of the cell culture room is generally that the aseptic operation area is located inside the room where the room is less moving, and the conventional operation and the closed culture are in a room, which is a clean room of a 100,000-level clean environment. The scrubbing and disinfecting work is located outside the clean room to avoid contamination of the indoor environment. The cell clean room requires an air conditioning system and maintains a certain positive pressure on the outside.
4, the anatomy laboratory is mainly used for the separation of organ tissues, need to have aseptic table, dissection microscope, various surgical equipment.
Second, laboratory equipment
2, carbon dioxide incubator
3, inverted microscope
4, pure water machine / ultra pure water machine
5, centrifuge
6, liquid nitrogen tank
7, low temperature refrigerator
8, the transfer window
9, autoclave
10, pipetting gun, electric pipetting gun
11. Various kinds of consumables required for cell culture, such as cell culture plates, cell culture bottles, and pipettes.
12, the rest of the instrument