The sterile room is usually a small room specially set up in the biological laboratory. The area should not be too large, about 4m2~5m2, and the height is about 2.5m. A buffer room, a purifying air shower room and a dressing room should be provided in the sterile outdoor room. Both the sterile room and the buffer room must be sealed. Indoor equipment must have an air filter. Ultraviolet lamps are installed in the sterile room and buffer room, and the ultraviolet light in the sterile room is lm from the work surface. The floor and walls in the sterile room must be flat, not easy to hide dirt, and easy to clean. The table top of the workbench must be level. Workers entering the sterile room should wear sterile clothes, caps, and shoes in the locker room, pass through the clean air shower, go to the buffer room, and then enter the sterile room for inspection.
Large-scale ultra-clean workbench is used in general laboratories in county-level cities. The ultra-clean workbench has a simple structure and is easy to move. There are purple lamps and purifying fans in the workbench. The outside is a push-pull small door or a glass door with two holes. When the operation is carried out, the arms can be inserted.
(1) The cleanliness of the aseptic operation room should reach 10,000 grades, the indoor temperature should be kept at 20 °C-24 °C, the humidity should be kept at 45%~60%, and the cleanliness of the clean bench should reach 100 grades. The number of colonies should be checked every month in the sterile room. In the state where the sterile room or the clean bench is opened, a number of sterile petri dishes with an inner diameter of 90 mm are taken, and the plate count agar melted and cooled to about 45 ° C is separately injected aseptically. 15 mL of the medium was placed and solidified, and then placed in a 36 ° C ± 1 ° C incubator for 48 hours. After the sterilization was confirmed, 3 to 5 plates were taken, placed in the left, middle, and right positions of the working position, and the cover was exposed. After a minute, it was placed in a 36 ° C ± 1 ° C incubator for 48 h, and taken out for inspection. The average number of plate bacteria in the 100-level clean area shall not exceed 1 colony, and the average clean room in Class 10000 shall not exceed 3 colonies. If the limit is exceeded, the sterile room shall be thoroughly disinfected until repeated inspections are satisfactory.
(2) The sterile room should be kept clean and tidy. Only necessary inspection tools such as alcohol lamps, alcohol cotton, lighters, tweezers, inoculation rings, scissors and glass pens should be stored indoors. It is strictly forbidden to pile up debris to prevent pollution.
(3) The sterile room should be provided with a working concentration of disinfectant, such as 5% cresol solution, 75% alcohol, 0.1% benzalkonium solution and so on.
(4) All instruments, medicines, glassware, etc. that need to be brought into the sterile room should be tightly packed or sterilized by appropriate methods. For example, the culture dish and the straw are sterilized in a dry box at 160°C-170°C for 1h~2h, and the physiological saline is sterilized at 121°C for 20 minutes by autoclave.
(5) The sterile room must be opened by UV lamp in the sterile room and buffer room for more than 30 minutes before use. At the same time, the purifying fan is turned on for ventilation. It is only half an hour after the UV lamp is turned off to enter the sterile room for work. After the sample is inspected, the remaining samples and various medicines and glassware should be cleaned in time, and all UV lamps should be turned on and sterilized for 30 minutes.
(6) The sample to be inspected should be kept intact before inspection and should not be opened to prevent pollution. Prior to testing, the outer surface was sterilized with 75% alcohol cotton balls and sterilized with ignited alcohol cotton if necessary.
(7) A blank control should be performed during each operation to check the reliability of the aseptic operation.
(8) When sucking the bacteria liquid, it must be sucked by the ear suction ball. Do not directly touch the straw with the mouth. If the straw touches the hand and other unsterilized objects, it must be replaced again to prevent pollution.
(9) Before and after each use, the inoculation needle must be sterilized by flame combustion, and the culture can be inoculated only after cooling.
(10) Pipettes, test tubes, etc. with bacterial liquid should be immersed in a disinfection bucket containing 5% of the solution of the sulphate solution, and taken out and rinsed in 24 hours; the culture dish should be re-sterilized in the autoclave. Handle the nutrient base, remember not to rush directly into the sewer to prevent blockage. If the bacteria liquid is sprinkled on the table or on the ground, it should be immediately overturned in the contaminated area for at least 30 minutes with 5% stone carbonate solution or 3% of the lysine, and then treated. If the work coat is contaminated by the bacteria liquid, it should be immediately Remove, wash after autoclaving.
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