1. ultra-clean bench?
At present, the vast majority of cellular laboratories use ultra-clean bench to achieve aseptic operation, with simple operation, easy to install, occupies a small space and purification effect is very good. Anhui people and purification for you to introduce the two main ultra-clean bench - lateral flow (vertical) and external flow (horizontal laminar flow). The working principle is generally the indoor air through the coarse filter initial filtration, by the centrifugal fan pressed into the static pressure box, and then by the high-efficiency air filter fine filtration, which sends out the clean airflow to a certain uniform cross-section of the wind speed through the sterile area, so as to form a high degree of cleanliness of the dust-free and bacteria-free working environment.
(1) Side-flow bench: air purified air flow from the left or right through the work surface to the opposite side, also from the top down or from the bottom up to the opposite side of the formation of airflow barriers to keep the work area sterile, the structure of the workbench for the closed type;?
(2) outflow bench: purified air flow toward the operator, so the outside air flow will not be mixed into the operation, the bench structure is open, but the operation of harmful substances experiments on the operator is not favorable. Ultra-clean bench should be required to regularly ask the relevant departments to check the cleanliness, meet the requirements of the ultra-clean bench cleanliness should reach 100, with a dust particle counter to detect the number of dust particles ≤ 5μm particle size should not be more than 3.5 / L; air flow should be controlled in the 0.75-1.0m3 / s; the number of bacterial colonies in an average of & lt; 1, according to the sterile condition of the need to replace filters when necessary.
2. microscope?
The inverted microscope is the daily work of the cell culture laboratory routine necessary equipment, mainly used for daily understanding of the growth of cells and observe the occurrence of pollution. If funds allow, it is recommended to choose the configuration of a photographic system of high-quality phase contrast microscope, anatomical microscope, fluorescence microscope, video system or time-lapse movie filming device, etc., can be photographed at any time and record cell growth.
3. Incubator?
Cells cultured in vitro, like cells in vivo, need to survive at a constant temperature, the general optimal growth temperature of 37 ℃, the temperature difference should not change more than ± 0.5 ℃. When the temperature rises 2℃, the change is unfavorable to the survival of cells, the temperature reaches 40℃ or more cells will die soon. Therefore, the constant temperature incubator and CO2 incubator which can precisely control the temperature are the best choice.
(1) Constant temperature incubator: should choose waterproof or transistorized self-control temperature incubator, such incubators have high sensitivity and more stable temperature control. General thermostatic incubator is cheaper, the disadvantage is that it is only suitable for closed culture.
(2) CO2 incubator: at present, most of the cell culture room has been widely used. CO2 incubator has the advantage of being able to provide a certain amount of CO2 (commonly used concentration of 5%), easy to stabilize the pH of the culture solution, suitable for open or semi-open culture. When using the culture bottle, in order to make the culture bottle and the outside world to maintain the ventilation state, can be slightly loosened bottle cap, in order to avoid cell contamination, the use of this type of culture, the incubator air must be kept clean, need to be regularly sterilized with ultraviolet radiation or alcohol, at the same time, the incubator should be placed in the tank containing sterile distilled water, to prevent evaporation of the culture solution, to maintain the relative humidity of the box at 100%.
(3) Cell culture consumables: the vessels for culturing cells can be used as Petri dishes, culture plates or culture flasks.
4. oven (drying oven)?
Some instruments and utensils used for cell culture must be dried before use, and glassware, etc. must be dry heat sterilized. Dry heat sterilization, the oven temperature should generally reach more than 160 ℃, usually using a blast oven. The advantage is that the temperature is uniform, the effect is better, the disadvantage is that the heating process is slow. Heating up can not be the first temperature after the blower and should be blown and warming at the same time to start, to 100 ℃, stop the blower. Need to pay attention to avoid wrapping the vessel of paper or cotton burnt, burnt debris can affect the growth of cells. Sterilization can not immediately open the door of the box, so as to avoid sudden cold lead to glassware rupture, should wait for the temperature to naturally drop to below 100 ℃ can open the door.
5. Water purification device?
Cell culture requires high quality of water, cell culture, cell culture-related liquid preparation of water as well as cleaning water for cell culture vessels must be strictly purified beforehand. At present, there are a variety of purification methods combined to purify ordinary water into pure water and ultra-pure water purification device is very flexible and convenient to use, can be wall-mounted, desktop, can be equipped with storage tanks, can also be used directly with the dispenser gun, but also according to the requirements of various types of experimental water can be configured to choose the function of sterilization, the effective removal of DNA enzymes, RNA enzymes, proteases, etc., and can effectively remove the heat, endotoxin, ultrafiltration-type water purification device.
6. Refrigerator?
Necessary equipment for cell culture laboratories, should be dedicated, shall not store volatile, easy to burn and other harmful substances to the cells, and should be kept clean. Generally include ordinary refrigerator, cryogenic refrigerator and ultra-low temperature refrigerator.
Ordinary refrigerator: store culture fluid, saline, Hanks liquid reagents and other items for culture, can be short-term preservation of tissue samples.?
-20 ℃ low-temperature refrigerator, ultra-low-temperature refrigerator: used to store the need for freezing to maintain biological activity as well as need to be stored for a long period of time preparations, such as enzymes, serum, etc.?
7. cell freezing storage?
Liquid nitrogen containers commonly used for storage are categorized into different types and specifications according to the need for use. Three factors need to be considered when choosing a liquid nitrogen container: volume size, ease of access and use, and liquid nitrogen volatilization. The size of liquid nitrogen containers is generally 25-500L, which can store about 250-15000 cryopreservation tubes of 1ml. The temperature of liquid nitrogen can be as low as -196℃, and care should be taken to avoid frostbite when using it. Since liquid nitrogen is volatile, it is necessary to pay attention to observe the amount of remaining liquid nitrogen and replenish it in time, so as to avoid cell damage or death due to insufficient liquid nitrogen. At present, there are many intelligent cell freezing storage devices to choose from, which can be configured with liquid nitrogen storage device with electronic controller to realize freezing and storage automation; and can monitor the liquid nitrogen level and sample temperature to ensure that the sample temperature is always at the set temperature point; it can be configured with an alarm system, which can be set up to alarm in case of malfunctioning of the liquid nitrogen level, temperature, battery, voltage, power supply, etc.; and it is equipped with hot gas bypass system to prevent warm air higher than -130℃ from entering into the liquid nitrogen tank, which can prevent cells from being damaged or died. It also has a hot gas bypass system to prevent the warm air above -130℃ from entering the liquid nitrogen tank, thus protecting the sample more effectively and preventing the container from heating up. In addition, you can choose the liquid nitrogen supply tank to replenish the storage tank with liquid nitrogen through the connecting pipe to ensure the safety of the sample.
8. Centrifuge?
Centrifuges are commonly used in cell culture to prepare cell suspensions, adjust cell density, wash and collect cells. The general routine configuration of 4000rpm tabletop centrifuge; if you want to do cell sedimentation, you need to use 80-100G centrifugal force, because too much centrifugal force will damage the cells. According to different requirements, there are large capacity, adjustable temperature centrifuges, high-speed centrifuges and cryogenic freezer centrifuges and other more functional centrifuges to choose from.
9. Balance?
Commonly used are precision balances and analytical balances. The accuracy of the analytical balance is generally 0.1mg, 0.01mg and 0.001mg. Generally, according to the amount of sampling and accuracy requirements to choose the right balance: sampling volume greater than 100mg should be used to select the accuracy of 0.1mg of the balance; sampling volume of 100-10mg, the use of precision 0.01mg of the balance; sampling volume of 10mg is suitable for the use of precision 0.001mg of the balance. The balance needs to be calibrated regularly, you can choose the balance with automatic calibration function, convenient maintenance. Balance use need to pay attention to clean, avoid corrosive powder, liquid direct contact with the weighing platform. Answer from