Tissue culture simulates the in vivo physiological environment in vitro under sterile, appropriate temperature and certain nutrient conditions. The tissue removed from the body survives, grows, and passes, and maintains its original structural and functional properties. Generalized tissue culture is synonymous with in vitro culture. Invitro cultures include all structural levels of culture, ie, tissue culture, cell culture, and organ culture.
Cell culture refers to the growth of cells including individual cells under in vitro conditions. The history of tissue culture has been nearly a hundred years. The initial tissue culture is an extension of embryology and microbiology. It is based on the principle of sterility. Natural body fluids (such as fetal juice, plasma) are used to maintain the tissue pieces cut from the whole. Observe the morphology and function of the cells.
The main difference between tissue cell culture technology and other general laboratory work is the requirement to maintain aseptic operation and avoid the effects of microorganisms and other harmful factors. At present, the extensive use of ultra-clean benches greatly facilitates tissue cell culture work and makes it possible for some routine laboratories to perform cell culture. Shanghai Creations offers: 60x15mm bacterial culture dish with lid, three outlets, product number: R01-66-1560-60x15mm?, price 374 yuan.
Cell culture laboratories should be able to perform six tasks: aseptic processing, incubation, preparation, cleaning, sterilization, and storage.
Sterile operation area
(1) Aseptic operation room:
The sterile operating area is limited to areas of cell culture and other aseptic operations, and is preferably isolated from the outside world, not accessible or otherwise interfered with.
The ideal aseptic operating room should be divided into three parts:
a) Dressing room - for changing clothes, shoes and wearing hats and masks.
b) Buffer room – located between the dressing room and the operating room, in order to ensure a sterile environment in the operating room, and to place a constant temperature incubator and some necessary small instruments.
c) 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. Shanghai Chuangsai provides: 9.0CM disposable cell culture dish, flat-bottom sterilization, suitable for cell adherent growth, commodity number: C81-TCD010090-10 / package, price 16.8 yuan.
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.
Air disinfection in aseptic operation room: UV lamp - produces ozone, and the indoor temperature and humidity are high, which is not conducive to the health of workers.
Air-filtered constant temperature and humidity device - best.
Clean room (zone) air cleanliness level table cleanliness level:
The maximum allowable number of dust particles / cubic meters 100 level 3,500 0 5 110000
The maximum allowable number of microorganisms is 350,000 2,000 100 3100000
Floating bacteria / cubic meters 3,500,000 20,000 500 10300000
Settling bacteria / dish 10,350,000 60,000 - 15
Ozone-free UV sterilizer electronic disinfection sterilizer--Under the action of high-voltage electric field, the internal and external electrodes of the electron tube undergo strong electron bombardment, which ionizes the air and converts the oxygen in the air into ozone. Ozone is a strong oxidant, which can react with the bacterial membrane and the enzyme protein hydrogen sulfide to oxidize and decompose, so that the diffuse diffusion of ozone gas can achieve the purpose of sterilization, and there is no dead angle when disinfecting. The residual ozone in the space after disinfection can be reduced to oxygen by itself for 30-40 minutes, the air does not leave odor, and the surface of the disinfected object is not left with residual poison.
(2) Purification workbench:
The operation is simple, the installation is convenient, the occupied space is small, and the purification effect is good.
There are two main types of purification workbench used in general bacterial culture rooms:
a) side flow or vertical
b) Outflow or horizontal laminar flow.
The working principle of the purification workbench (substantially the same)--the air is usually filtered through the coarse filter, and the centrifugal fan is pressed into the static pressure box, and then filtered by the high-efficiency air filter to clean the airflow. A uniform cross-section wind speed passes through the sterile field to create a dust-free, sterile, high-clean work environment.
Side-flow workbench—The airflow after air purification flows from the left or right side through the work surface to the opposite side, and also from the top to the bottom or from the bottom to the opposite side, forming an airflow barrier to keep the working area sterile. The workbench structure is closed. Outflow (horizontal) workbench—The purified air flows toward the operator, so the external airflow does not mix into the operation, but the hazardous substance test operation is unfavorable to the operator. The workbench structure is open (already used).
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