Standard Operating Procedures for Biosafety of Cell Culture Room II
Part One: General Requirements for Biosafety of Cell Culture Room II
1. Purpose
In order to standardize the safe operation of microorganisms in the cell culture room II, to ensure laboratory biosafety, to ensure the safety of laboratory personnel and the laboratory environment from contamination.
2. Scope of application
It is suitable for the safe operation process of the central cell culture room II.
3. Requirements
3.1 Experimenters who enter the laboratory to carry out experimental projects should first fill in the "Application Form for Cell Culture Room II of the Second Hospital Clinical Research Center". Relevant training will be carried out after the approval of the instructor and the center, and the training can be carried out only after passing the training.
3.2 Each experiment must be booked in advance.
3.3 It is forbidden to engage in activities unrelated to the experiment in the laboratory, and the door of the laboratory should always be closed.
3.4 The staff shall not keep long nails, and shall not wear rings, bracelets or other accessories that may reduce the protective effect of protective equipment.
3.5 It is forbidden to store non-experimental items or reserve experimental items that will not be used for a long time in the laboratory.
3.6 The type of virus that can be operated in this laboratory is a carrier virus (in line with the BSL-2 standard), and all pathogenic microorganism operations cannot be carried out in this laboratory.
3.7 Personal protection
Personal protective equipment must be used when working in the laboratory.
Personal clothes and ordinary laboratory clothes cannot be worn into the working area of the cell culture room II.
Long skirts, short skirts and shorts are not allowed in the laboratory; open-toed shoes are not allowed in the laboratory.
It is strictly forbidden to handle and wear glasses in the laboratory.
It is strictly forbidden to leave the laboratory wearing special protective clothing for cell culture room II.
Damaged gloves should be replaced immediately.
3.8 Operating specifications
The cell culture II room includes a buffer zone and a core operating area. Only contaminated materials sealed in sterilized containers can be brought to the buffer zone; viruses can be manipulated in the core operating area.
Any operations with active viruses should be carried out in a Class II biological safety cabinet. The open experiment platform does not allow operation of such viruses.
The laboratory should be kept clean and tidy. Only the instruments and objects currently in use can be placed on the laboratory bench. Consumables should be stored in designated boxes and picked up as needed.
Syringes and needles should only be used when absolutely necessary. The used needles must be placed in a special sharps box.
Plastic equipment should be used, and glass products can only be used when needed in the experimental procedure (for example, using a microscope or phenol slide).
Must use filter tips and straws.
All tubes, bottles, culture plates, etc. must be clearly marked, and can only be placed in the incubator, refrigerator or freezer in the cell culture II room.
Active viruses are not allowed to be taken out of the cell culture room II unless approved by the person in charge of the laboratory (for example, for the purpose of transporting specimens).
Under no circumstances are books and laboratory notebooks allowed to be brought into the Cell Lab II.
Part Two: Guidelines for the Entry and Exit of Cell Culture Room II
1. Purpose
Formulate standard operating procedures for personnel entering and exiting the cell culture room II to ensure laboratory biosafety.
2. Scope of application
It is suitable for personnel entering and exiting the cell culture room II.
3. Regulation requirements
3.1 Enter the program
Put on the shoe covers before entering the buffer room, then wear a hat, mask, and gloves (the cuffs of the work clothes should be tied inside the gloves);
When entering the workshop to operate, you must wear protective clothing to operate. Another layer of gloves can be worn (the cuffs of the protective clothing should also be tied inside the gloves).
3.2 Exit the program
When exiting the workshop, take off the outermost gloves, put them inside out, and put them in an autoclave bag; take off the protective clothing, hang it in the designated position, and disinfect once a week; if the protective clothing is contaminated by virus fluid, replace it immediately New protective clothing, wipe the contaminated area with 10% disinfectant, and then put it in an autoclave bag; take off the cap and mask, and put it in the autoclave bag.
When exiting the buffer room, take off the shoe covers and inner gloves and put them in the designated garbage bag; close the entrance door of the cell culture room II.
Part Three Standard Procedures for the Use of Cell Culture Room II
1. Purpose
This procedure is specially formulated to ensure the sterility of the cell culture room II environment.
2. Scope of application
It is suitable for the monitoring of air microorganisms in sterile rooms.
3. Process requirements
Before starting work, UV irradiation in the workplace and the biological safety cabinet for 30 minutes, and then the biological safety cabinet should be operated for at least 15 minutes.
First wipe the walls and countertops of the biological safety cabinet with 70% alcohol, and then put the experimental supplies in the cabinet.
Only the materials and supplies necessary for this experiment are placed in the biological safety cabinet. In order to avoid cross-contamination, the laboratory supplies should be disinfected with 70% alcohol before being put into the biological safety cabinet.
At the end of the experiment, all items in the biological safety cabinet, including equipment, samples, and reagents, should be cleaned of surface contamination, and then the items should be removed from the safety cabinet.
After the work is over, use 70% alcohol to wipe the walls and countertops of the biological safety cabinet. Ultraviolet radiation disinfection for 1 hour (automatically cut off after 1 hour) to strengthen the disinfection effect. At the same time, ultraviolet irradiation disinfection in the workplace for 1 hour.
The sterility inspection record shall be carried out once a month in the sterile room.
Part IV: Standard Practice for Disinfection of Cell Culture Room II
1. Purpose
Protect the biological safety of the cell culture room II.
2. Scope of application
Daily cleaning of the laboratory, disinfection of polluted waste generated during the experiment, used protective equipment and facilities and equipment.
3. Process requirements
The laboratory staff should take good personal protection. Wear work clothes, gloves, masks, and hats.
Preventive disinfection: wipe the object to be disinfected with 70% alcohol for more than 15 minutes.
Spill contamination: Cover and absorb the spill with absorbent paper towels, pour an appropriate 10% disinfectant (effective chlorine concentration of 5000mg/L) on the paper towels to cover the surrounding area. When using disinfectant, start from the periphery of the overflow area and work towards the center. The action time is more than 30 minutes.
During the operation, all pipettes, Plastic transfer pipettes and the inner and outer walls of the pipette tips that have been exposed to the virus must be treated with 10% disinfectant before throwing them into the garbage collector;
Please put the paper sleeve of the pipette and the jacket of the transfer pipet as solid garbage in the garbage bag and spray 10% disinfectant into the garbage bag.
After the experiment, put all the garbage in garbage bags, and spray 10% disinfectant into the garbage bags.
Autoclave the solid waste twice a week.
