Lead Is your cell culture laboratory a well-functioning cell culture laboratory? It is not only reasonable in layout, but also safe and convenient to operate, and can avoid pollution. It can not only meet the needs of work, but also ensure that the test results are not disturbed. The fireproof, moisture-proof and anti-corrosion properties are all available, and the functions of ventilation, purification, disinfection and sterilization are essential. By following the principles of safety, efficiency, comfort and “people-orientedâ€, we can build a standardized and standardized cell culture laboratory to achieve the best use efficiency. First, cell culture laboratory design and layout The research on tissue cell culture technology is not general. It is a biological unit with complete vitality. Therefore, unlike other general experimental work, it is required to maintain aseptic operation and avoid the influence of microorganisms and other harmful factors. Cell culture laboratories should be able to perform six tasks: aseptic processing, incubation, preparation, cleaning, sterilization, and storage. It is best to set up each department separately in the connected rooms. In particular, the aseptic operation room can be set separately. If they are placed in a large laboratory, the aseptic operation area and the cleaning and disinfection area should be located at both ends. The preparation, storage and incubation zones are located between the two zones. Aseptic operation area (1) Aseptic operation room: It is only limited to the area of ​​cell culture and other aseptic operations. It is best to be isolated from the outside world, not to walk through or interfere with other disturbances. The ideal aseptic operating room should be divided into three parts: • Dressing room – for changing clothes, shoes and wearing hats and masks. • Buffer room – located between the dressing room and the operating room, in order to ensure a sterile environment in the operating room, while placing a constant temperature incubator and some necessary small instruments. • Aseptic operation room – dedicated to aseptic processing, cell culture. The size should be appropriate, and the top should not be too high (not more than 2.5m) to ensure the effective sterilization effect of ultraviolet rays; the wall is smooth and has no dead ends for cleaning and disinfection. The workbench should not be placed against the wall. The countertop should be smooth pressed and molded into a surface, painted white or gray to facilitate the observation of the anatomical structure and phenol red. (2) Ultra-clean workbench/biological safety cabinet: easy to operate, easy to install, small footprint and good purification effect. The basic principle of both has the process of passing the gas through the high-efficiency filter to form a dust-free and sterile high-purity target environment. Therefore, the performance of the fan and high-efficiency filter configured by the instrument directly determines the target environment. Degree of degree. The difference between a clean bench and a biosafety cabinet: 2. Incubation area Although the requirements for sterility in this area are not stricter than the aseptic area, they still need to be clean and dust-free, so they should also be placed in areas where there is less interference than walking through. In general, the laboratory uses incubators for incubation. 3. Preparation area In this zone, preparation of a culture solution, a liquid for culture, and the like is mainly performed. After the preparation is completed under the conditions of balance, P H meter, magnetic stirrer, etc., it should be filtered and sterilized at the aseptic workstation. Liquid preparation seems simple, but it is also sloppy. It is directly related to the success or failure of tissue cell culture. If it is not strictly aseptic, it may stage the annual drama - blue thin mushrooms. 4. Storage area The requirements for the storage area are convenient for picking and placing, mainly including refrigerators, drying boxes, liquid nitrogen tanks, sterile culture liquids, culture bottles, etc., and the environment also needs to be clean and dust-free. 5. Cleaning and disinfection area The cleaning and disinfection area should be separated from other areas, mainly for cleaning, preparation, disinfection and preparation of three steamed water for all cell culture vessels. Second, commonly used basic equipment Tissue cell culture research and production of necessary equipment, lacking can not work. Instrument (1) Microscope In order to grasp the growth of cells and observe the presence or absence of pollution. If the laboratory funds permit, you can configure a high-quality phase contrast microscope with a photographic system, you can photograph and record the situation of the cells at any time, which is very helpful for the development of scientific research. If conditions permit, you can also add a dissecting microscope, a fluorescence microscope, Video system device. (2) Incubator Cells cultured in vitro, like cells in vivo, need to survive at a constant and appropriate temperature. The optimum culture temperature for most mammalian cells is 35 to 37 ° C. When the temperature is raised by 2 ° C, the cells are not tolerated for several hours, and the temperature above 40 ° C will soon die. Therefore, an incubator capable of controlling the temperature is required. Such as constant temperature incubators and carbon dioxide incubators, have higher sensitivity. In addition, most animal cells need to survive and grow in a slightly alkaline environment, with a pH of 7.2 to 7.4, and no more than 6.8 to 7.6. CO 2 is both a desired component of cells and a cellular metabolite. It forms a buffer system with sodium bicarbonate in most media to maintain physiological pH. The carbon dioxide incubator can precisely control the concentration of carbon dioxide, but the ordinary constant temperature incubator cannot. Carbon dioxide incubators are often used in cell culture laboratories that are now open or semi-open. Widely recognized by researchers is a gas-encased, carbon dioxide incubator equipped with infrared sensors. These incubators are more sensitive than ordinary ones, and the temperature and carbon dioxide concentration are also stable. (3) Oven Some instruments and utensils used for cell culture should be dried before use. Glassware needs to be dry and heat sterilized. Therefore, the cell culture laboratory is equipped with an electric drying oven/oven. (4) Water purification device Cell culture requires high quality water, and the water used must be purified first. An ion exchange pure water device or a distiller can be used for water purification. (5) refrigerator The cell culture room must be equipped with a regular refrigerator or freezer, and the refrigerator preferably has a low temperature refrigerator (-20 ° C). The former is used for storing culture materials such as culture fluids and physiological saline, and for storing tissue specimens for a short period of time. The -20 ° C ultra-low temperature refrigerator can be used to store preparations that need to be frozen to maintain biological activity and stored for a longer period of time. (6) Cell freezing storage Cell culture work often requires storage of cells, and the temperature of liquid nitrogen is very low, reaching -196 ° C. At this temperature, the material metabolism and life activities of the cells are almost completely stopped. Therefore, liquid nitrogen containers are commonly used to store cells. Prevent frostbite when using. As the liquid nitrogen is continuously volatilized, attention should be paid to the observation of the residual liquid nitrogen, and the liquid nitrogen should be replenished regularly to avoid excessive evaporation and damage to the cells. (7) Centrifuge Used to prepare cell suspensions, adjust cell density, wash, and collect cells. (8) sterilizer Items that are in direct or indirect contact with cells need to be sterilized. If there is a uniform disinfection supply room, some commonly used, larger items can be provided after the room is sterilized and sterilized. Even so, sometimes the items that need to be added temporarily or the fragile utensils still need to be disinfected by the cell culture laboratory. Therefore, the sterilizer of this laboratory should be equipped, and the high pressure steam sterilizer is often used. (9) Balance and filter The reagents required for culturing the culture medium, the trypsin solution for digestion, the buffer solution, etc. need to be weighed with a balance, and after being configured, it is still filtered and sterilized by a filter before being used for cell culture. 2. Culture utensils Requirements: good transparency, non-toxic Vessels related to culture operations: pipettes, pipettes In addition, centrifuge tubes for collecting cells, tubes for placing reagents or temporarily inserting pipettes, and blood cell counting plates or automated cell counting instruments for cell counting are all equipment that the cell culture laboratory needs to configure. Sometimes it is necessary to store small pieces of cultured items. For the convenience of high temperature sterilization, many laboratories choose to use aluminum lunch boxes for contents. Third, special equipment In order to carry out cell science research more efficiently, accurately and deeply, conditional (local tyrants) laboratories can add special, advanced and sophisticated equipment such as multi-functional enzyme-linked immunosorbent analyzer, fluorescence microscope, flow A cytometer, a cell expansion system (bioreactor), and the like. Bioreactor Conclusion Esco's sharing of standard cell culture laboratory construction allows you to face the cell culture laboratory construction or upgrade and be willing to say So easy! Xi'an Double H Health Technology Co., Ltd , https://www.dhextract.com
