Safety

Working in a Level2 Physical Protection Laboratory
Each member of our team receives lab training to understand the safety regulations required to work in the PC2 lab. Upon entering the lab, all team members were mentored by our lab supervisor, Dr. Li, for three weeks to ensure safe operation and familiarization with the lab. During subsequent lab operations, we ensured our personal safety through protective gear, sterilization of ourselves and our space, proper waste disposal and sealing of biological components. We learned that our PC2 lab is at Bio-safety Level 2 and requires moderate controls. The nature of our program requires the generation of many genetically engineered organisms and we are careful to handle them properly.
 
Image 1 Laboratory garbage to implement wet and dry separation system; organic toxic reagents after use should be poured into the waste liquid barrel to wait for centralized treatment, the waste liquid barrel must be placed under the corrosion-resistant tray to prevent leakage of waste liquids; solid waste should be placed in the medical waste trash cans to wait for centralized treatment, and medical waste waste cans should be placed in a special area (as shown in the Image of the yellow line).

Use of Organisms and Reagents
We used BL21 DE3 E. coli throughout this project. Before using any bacteria, we carefully researched their potential risks to understand the necessary level of caution. The bacteria we constructed were not dangerous inside or outside of our project. Even so, we took great care in handling any biological products, using bleach to kill any bacterial waste and treating contaminated materials as autoclaved.

Image 2 The autoclave must be trained and operated according to the instructions on the wall when in use; the autoclave must be guarded next to the autoclave when it is in operation; and it must be opened only when the temperature is reduced to less than 60°C and the internal pressure is 0 at the end of use.

We use several hazardous reagents in our projects. We took extra precautions to keep ourselves safe by carrying out an initial risk assessment and wearing extra safety equipment for handling.

To test the activity of metal-binding proteins, we used an aqueous solution of inorganic cadmium (0.05 mol/mL). We use this highly toxic substance only after a thorough risk assessment. In the laboratory, our safety measures include the use of specialized chemical storage cabinets with cytotoxic fume hoods dedicated to the handling of cadmium. We also use rugged nitrile gloves and safety goggles over standard personal protective equipment to prevent harmful chemical exposure.
 
Image 3 Aseptic experiments must be performed in an ultra-clean table, with lab coats and hair tied up, masks and sterile gloves, and all items in the ultra-clean table must be sterilized and an alcohol lamp lit to create a sterile circle.
 
Image 4 Hazardous chemical storage cabinets; a strict two-person, two-lock registration system is in place to ensure the safe use of chemicals.
When testing for protein expression, we handled sodium dodecyl sulfate (SDS). From our risk assessment, we learned that SDS is an extreme irritant. To minimize this risk, we wore masks and goggles while handling SDS to prevent inhalation and possible eye irritation.

 

Installations
In our lab orientation, we learned about the possible hazards of lab equipment, including ultrasounds, safety cabinets, and centrifuges. Since this equipment was necessary for our experiments, we took extra steps to ensure our safety. When using the sonicator, we wore protective headphones to minimize the risk of ear damage. We made sure to check the contents of our fume cupboards and bio-safety cabinets prior to use as there could be harmful chemicals and biologicals present. Our training in the use of centrifuges means we understand how to balance the machine correctly to prevent potential bodily harm.
 
Image 5 High-pressure gas cylinders must be stored in sealed explosion-proof cabinets, with a two-person, two-lock system; cylinders should be relieved of pressure when not in use, and the sealing status of cylinders should be checked regularly.

 

The following is the Code of the State Key Laboratory of Bioelectronics at Southeast University

I. Laboratory Safety and Management
1.1 General Principles
1 The laboratory should be kept serious, quiet, neat and clean, smoking is prohibited, and the introduction of people and objects unrelated to the experiment is not allowed.
2 When doing experiments must be dressed in overalls, experiments should be conscientious and responsible, to comply with the operating procedures, experiments in the middle of the workplace may not leave without authorization.
3 Pre-use of unfamiliar items should be preceded by an understanding of their properties, which can be obtained through the Material Safety Date Sheets (http://www.msds.com/). Whenever flammable and explosive substances are used or toxic, hazardous and infectious substances are tested, they should be agreed by the relevant management personnel of the laboratory and recorded, and all laboratory personnel should be informed at the same time. It should also be operated in the designated location as indicated, and appropriate reprocessing should be carried out after the experiment is completed.
4 Precision and valuable instruments should be kept and used by specialized personnel, and the registration procedures for their use should be strictly implemented; damage to or malfunctioning of instruments should be reported immediately to the relevant personnel so that they can be dealt with as appropriate.
5 Laboratory use is completed, must be cleared in a timely manner; experiments after the instrument should be cleaned up in a timely manner, reagents should be returned to the original place in a timely manner, to maintain the tidiness of the laboratory.
6 The access card used in this laboratory is for personal use only. Loss of the card must be reported promptly and a new card must be issued.

1.2 Laboratory Routines
1 All personnel engaged in laboratory operations must be trained and qualified by the staff of the room before entering the laboratory for duty. New faculty and students should enter the laboratory to carry out experimental work under the guidance and training of other faculty and students in the room.
2 The personnel entering the laboratory should strictly observe the routine operating procedures of the laboratory.
3 When using or testing toxic, hazardous and infectious substances, they should be operated in the designated location and clearly marked to indicate what kind of operation is being carried out or will be carried out continuously, and the corresponding post-processing should be carried out after the completion of the experiment; strong acids and other corrosive and irritating reagents should be opened for use in a fume cupboard, and the remaining reagents should be processed accordingly.
4 laboratory precision instruments are used, must be cleared in a timely manner, and make use of the registration.
5 All reagents, appliances used, must be put back to the original place, the experimental table and the experimental cabinet reagents, appliances, in order to prevent cross-contamination, do not use at will.
6 Comply with laboratory safety measures and familiarize yourself with facilities such as alarms, smoke evacuation, fire fighting, accident lighting, etc., the use of corrosive, explosive, flammable and toxic substances and the corresponding safety measures.
7 The last person to leave the laboratory is required to check that all labeled instruments and power supplies are turned off.

1.3 Incident Analysis and Reporting System
1 Accident in this system means:
1.1 Anyone who damages precision instruments and valuable utensils resulting in direct and indirect economic losses of 500 yuan or more.
1.2 Anyone who violates the operating procedures, neglects his/her duties and causes combustion, explosion, workplace accidents or serious losses.
1.3 Those who have caused serious consequences or economic losses due to the disclosure of test data or technical information.
1.4 Loss of confidential documents and technical files.
1.5 Failure to maintain water, electricity and gas equipment in a timely manner leads to serious consequences.
1.6 Any other negligence resulting in serious consequences.
1.7 A person who has caused illness or injury to another laboratory member as a result of a laboratory accident or omission.
2 After the accident, timely measures should be taken to prevent the accident from expanding and to minimize and eliminate the effects of the accident.
3 When an accident occurs, it should be reported to the relevant personnel in a timely manner so that it can be dealt with as appropriate, and should not be withheld, or else the treatment will be aggravated.
4 Those responsible for major accidents shall be held financially and legally liable, depending on the circumstances.
Attachment: Contents of "Accident Report Form"
1 Overview of the Incident: time, place, persons involved and witnesses
2 Brief description of the incident
3 Accident losses (including direct and indirect economic losses, treatment costs, impact, etc.)
4 Remedial and improvement measures taken
5 Key lessons
6 Disposition.

1.4 Instrumentation and Reagent Management System
1 Instrumentation acceptance, installation and commissioning should be completed after the production certificate of conformity, instruction manual and other complete information should be organized and filed for preservation.
2 All instruments and equipment should be identified for specialized storage, regular inspection, maintenance and repair. General instruments and equipment, the use of personnel must be proficient in the operating procedures, beginners should be skilled in the operating procedures of the instrument under the guidance of the operation of personnel. Valuable instruments should be operated by specialized personnel.
3 Anyone must be used in accordance with the operating procedures of the instrument, if there is any abnormality, you must immediately stop using and report to the relevant personnel. Only after troubleshooting will it be allowed to be used, and it is strictly prohibited to work with faults.
4 No outside personnel will be allowed to operate the laboratory instruments alone.
5 General reagents are neatly arranged in reagent cabinets and organized periodically as needed.
6 Self-prepared reagents need to be labeled on the reagent bottle with the name of the reagent, the person who prepared it, and the date of preparation.
7 Toxic and hazardous reagents must be fully and clearly labeled and marked and placed in a safe and secure place.

1.5 Laboratory Hygiene and Office Systems
1 Laboratory health by temporary workers to arrange cleaning, other personnel should respect the labor of others, abide by social morality, and consciously maintain the environmental health of the laboratory.
2 Visitors to the laboratory must be accompanied by the relevant laboratory personnel before entering the laboratory.
3 The laboratory configuration of the computer should be used for work and study, work time is not allowed to play games, watching movies, online chatting, and strive to improve the efficiency and quality of work.

1.6 Research Records and Retention System
1 All research work is recorded in the prescribed research log book.
2 The content of the record should be true, systematic, neat and original, mainly including the date of the experiment, temperature and humidity, experimental methods, operation steps, the name of the instruments and equipment used, experimental results, data processing, results analysis, discussion and so on.
3 The scientific research record book is an integral part of the scientific and technological archives and an important basis for publishing papers and declaring achievements. The page number of the original scientific research record book must be indicated in the application form for proof of publication.
4  Students are required to submit a lab notebook for graduation.