1. Common equipment
1. Equipment in preparation room
Single distilled water distiller, double distilled water distiller, acid tank, oven, pressure cooker, storage cabinet (storing unsterilized articles), storage cabinet (storing sterilized articles), packaging table. Equipment in the solution preparation room: torsion balance and electronic balance (weighing medicine), PH meter (measuring PH value of culture solution), magnetic stirrer (configuring solution room to stir solution).
2. Equipment of culture room
Liquid nitrogen tank, storage cabinet (storing sundries), fluorescent lamp and ultraviolet lamp, air purifier system, low temperature refrigerator (- 80 ℃), air conditioner, carbon dioxide cylinder, side table (writing test records).
3. Equipment that must be placed in the sterile room
Centrifuge (collecting cells), ultra-clean worktable, inverted microscope, CO2 incubator (incubating culture), water bath, three-oxygen disinfection and sterilization machine, 4 ℃ refrigerator (placing serum and culture solution).
2、 Aseptic operation
（1） Sterilization of sterile room
1. Clean the sterile room regularly: once a week, first use tap water to mop the floor, wipe the table, and clean the working table, and then use 3 ‰ lysol or bromogeramine or 0.5% peracetic acid to wipe.
2. Sterilization of CO2 incubator (incubator): first wipe with 3 ‰ bromogeramine, then wipe with 75% alcohol or 0.5% peracetic acid, and then irradiate with ultraviolet lamp.
3. Sterilization before experiment: turn on the ultraviolet lamp, the three-oxygen sterilizer and the air purifier system for 20-30 minutes respectively.
4. Sterilization after the experiment: wipe the ultra-clean table, side table and inverted microscope stage with 75% alcohol (3 ‰ bromogeramine).
Sterilization preparation of laboratory personnel
1. Wash hands with soap.
2. Wear isolation clothes, isolation caps, masks and slippers.
3. Wipe hands with 75% alcohol cotton ball.
Demonstration of sterile operation
1. All bottles of alcohol, PBS, culture medium and trypsin brought into the ultra-clean workbench should be wiped with 75% alcohol on the outer surface of the bottle.
2. Operate near the flame of alcohol lamp.
3. Utensils must be sterilized before use.
4. The utensils (such as bottle caps and droppers) that continue to be used should be placed at a high place, and should still be overheated during use.
5. All operations should be close to the alcohol lamp, and the action should be light and accurate, and should not be touched randomly. If the straw cannot touch the waste liquid tank.
6. When aspirating more than two kinds of liquid, pay attention to replacing the suction pipe to prevent cross contamination.
See the next chapter for disinfection of instruments.
Post time: Feb-01-2023