Difference between revisions of "Team:CSMU Taiwan/Safety"
| Line 277: | Line 277: | ||
</div> | </div> | ||
<div class="col-md-9"> | <div class="col-md-9"> | ||
| − | <h3> | + | <h3>Lab safety</h3> |
| − | + | <br> | |
| − | + | <p> Experiments regarding proteins of our iGEM project were conducted in the Biosafety Level-1 laboratories, whereas experiments associated with virus were conducted in the Biosafety Level-2 Laboratories. Both labs are under the regulation of Environmental Safety and Health Center of Chung Shan Medical University. The emergency exits and basic risk assessment are the first things to check every day prior to experiments.</p> | |
| + | <p><b>Training</b></p> | ||
| + | <p> All members of the laboratory were well-trained by the mentors and had ability to conduct experiments in a safe environment. People who entered the laboratory should follow the established laboratory safety principles of our school. Master’s degree students of the lab and our instructor helped us to complete the safety training.</p> | ||
| + | <p style="color:#009698">Special courses</p> | ||
| + | <p style="color:#009698"><b>Conference for Lab Safety, Microscope and Common Equipment Usage held by Department of Biomedical Sciences</b><p> | ||
| + | <p>By attending the conference, we learned several important issues regarding lab safety as listed below:</p> | ||
| + | <p><b>1. Safety precautions and operating notice of commonly used instruments:</b><p> | ||
| + | <p><li>Freeze Dryer</li></p> | ||
| + | <p>The order to start the machine is of great importance. First, we should turn on the machine in the middle to warm it up for about 30-60 minutes. Then we turn the pump open. Note that if any water drop slides into the liquid, the pump will rise. Therefore, remember to drop the water droplets slowly from below.</p> | ||
| + | <p><li>Low Temperature High Speed Centrifuge</li></p> | ||
| + | <p>Balance is very important! Balance the weight before putting the samples into the machine. Do not mess with the tools provided by a different company from the machine manufacturer. For example, if we want to centrifuge with 15 cc centrifuge tube, we use the centrifugal sleeve provided by the manufacturer instead of plugging the tube into any other centrifugal sleeve. Under high speed centrifugation, unbalance could cause great damage to the middle shaft of the machine (which is the heart of it), and would be very dangerous to operators since the samples could burst out from the machine in a very high speed. Don’t put any thing on the machine when it is operating as well.</p> | ||
| + | <p><li>Laminar Flow (Hood)</li></p> | ||
| + | <p>The flow should be turned on for at least 20 mins before usage. The hood must be pulled down when the UV lights are on</p> | ||
| + | <p><li>Autoclave</li></p> | ||
| + | <p>Don’t put any PC materials into the machine or it could melt. Always remember to check if both the drainage knob and exhaust knob has been tightly closed before powering up the machine. Ensure that the door is fully sealed, however, don’t over-tighten the bolt as it may result in damage to the gasket. Before taking out the materials, turn the exhaust knobs loose. Gloves are necessary.</p> | ||
| + | <p><li>Ultrasonic washer</li></p> | ||
| + | <p>The volume of water column needs to be adjusted according to the black indicator line. The water will drain out if it exceeds the line .</p> | ||
| + | <p><li>Micro scales</li></p> | ||
| + | <p>Items must be placed in the middle of the scale. Always check the gradienter to see if the scale is standing at a horizontal platform .</p> | ||
| + | <br> | ||
| + | <p><b>2. School institutions that we can consult with when facing questions with the safety of experiments</b></p> | ||
| + | <p>˙ Always feel free to ask for help at the Environmental Safety and Health Center! If there is any problem regarding the instruments, please call the department for help, and the department will contact the company to handle it.</p> | ||
| + | <p><b>3. Other suggestions for improving the safety of laboratory operations:</b></p> | ||
| + | <p>If you’re uncertain about any step during the experiment, STOP! Never feel embarrassed to ask for help !!</p> | ||
| + | <br> | ||
| + | <p><b>MSDS</b></p> | ||
| + | <p>Material Safety Data Sheet, MSDS. Every chemical, reagents in the lab are printed and collected in this data folder. The folder is placed in a significant place near the door, which is easy to find and read in case of emergency.</p> | ||
| + | <br> | ||
| + | <p><b>Biohazard waste Management</b></p> | ||
| + | <p>All kinds of waste produced in the working area of the laboratory are considered biohazard waste, including every experiment and consumptive material. Those biohazard wastes would be collected into a specially labelled red plastic bag, and be sent to the Environmental Safety and Health Center of Chung Shan Medical University for further disposure process.</p> | ||
<!----Empìezo listado con una separación----> | <!----Empìezo listado con una separación----> | ||
<table style="width:100%"> | <table style="width:100%"> | ||
Revision as of 07:03, 14 October 2019
Introduction
The ultimate aim of technology is to create a better world for human beings. However, risks arise during the development of these techniques, varying from physical harm to privacy invasion. The advantages of technology can never be fully demonstrated if these potential hazards presents, therefore, safety should be of the top priority no matter for researchers or users.
Our team aims to develop a more essential influenza rapid test tool. Meanwhile, creation of a new diagnostic tool takes more than research and development. Implementation of such a device means it will be brought into direct contact with society. To ensure that our work is safe for the end user and to prevent miscommunication about the purpose of the technology, it is also essential to consider the social aspects that our tool will bring.
To sum up, in the safety section, we are going to explore four different aspects of our project, respectively Biosafety, Lab Safety, Regulations, and Ethics.
Biosafety
Labsafety
Regulations
Ethics
Biosafety
Our team complies with the third edition of Laboratory Biosafety Manual released by World Health Organization.
Moreover, according to Taiwan Centers for Disease Control(Taiwan CDC), some of the current status for Biosafety are stated as bellow:
In 2018, Taiwan CDC completed editing, revision, and announcement of the “Guideline for Governing Management of Select Agents and Toxins”, “Guidance on the Inventory of Select Agents and Toxins”, “Security Guidance for Select Agent or Toxin Facilities”, “Guidance for Suitability Assessments”, “Drills and Exercise Guidance”, and “Select Agents or Toxins Biosafety/ Biocontainment Plan Guidance”. We also accomplished the amendment of the “Regulations Governing Management of Infectious Biological Materials” on January 31, 2019, categorizing select agents and toxins as a special section.
What our team did to comply with it
Although no select agents or toxins on the list are involved in our experiment, we still looked through the guidance carefully.
Nucleoprotein and hemagglutinin of Influenza A and Influenza B virus are our target protein for SELEX, meanwhile whole influenza viruses are used to test the inhibition ability of our selected aptamers. In addition, in the Charactization and Improvement section, we improved the water solubility of HA1(Hemagglutinin for H1N1). The subtypes we chose are classified as seasonal influenza, which are surprisingly potential select agents too.
Apart from complying to the Biosafety manual for Level 2 labs, we also took some effort looking through Dual Use Research of Concern, DURC. After discussing with our instructor, we drafted a Risk mitigation plan.
2. Biosafety Mechanism Registration
By December 2018, 502 entities reported biosafety management units to Taiwan CDC, of which 496 entities established biosafety committees and 6 entities designated a biosafety specialist staff. These included 20 government agencies, 153 medical institutions, 54 academic research institutions and 275 other groups.
What our team did to comply with it
We are proud to announce that our institution, Chung-Shan Medical University, is one of the registered entity! We have applied for agreement to use virus in our experiment at the safety committee of CSMU, and the safety committee checked our facility, gave us a quiz regarding associated operations before permitting us. In addition, CDC tracks our usage regarding all contagious agents including influenza virus.
3. Biosafety Inspections of High-Containment Laboratories
Since 2006, Taiwan CDC has routinely inspected BSL-3 and above laboratories to monitor operations and ensure safety. In 2009, TB-containment laboratories, which process culture manipulation for identification and drug-susceptibility tests, were added to inspections. In 2019, Taiwan CDC completed the annual routine laboratory biosafety and biosecurity inspecting of 22 high-containment laboratories (refers to Level 3 and Level 4 biosafety and animal biosafety laboratories.) and RG3 pathogens storage facilities.
In addition, Taiwan CDC supervised the local government health bureaus to complete the laboratory inspecting of 75 BSL-2 microbiology laboratories of biotechnology industries. In order to strengthen biosafety and biosecurity of the entities that possess or use select agents and toxins, Taiwan CDC has counselled and visited the facilities of 7 entities in 2018.
What our team did to comply with it
All of our experiments regarding virus were conducted in the Biosafety Level-2 laboratories. Under the supervision of CDC, our instructor manages the lab strictly and obeys to all guidance written in the manual. For example, we were never allowed to enter the lab before putting on gloves, masks and lab coats. The wastes were always carefully collected and dispensed. Most importantly, our instructor seldom leaves us alone when performing experiments that could infect disease accidentally.
4. Laboratory Biosafety Education and Training
In 2018, 4 e-learning courses on laboratory biosafety were recorded. Taiwan CDC also organized 3 biorisk management and 3 select agents and toxins training course sessions, with a total attendance of 429 laboratory workers. Taiwan CDC developed “Laboratory biosafety management information system” (ver. 2.0) which helps to conduct and oversee the condition of management of storage and transformation of infectious biological material. We held 10 operation training and 2 systematic data maintenance symposiums to help all entities getting familiar with the system, and 508 laboratory workers attended.
What our team did to comply with it
We have organized to watch these videos when facing the same situations the videos focused on.
5. Laboratory Biorisk Management
In 2018, the bio-risk management system using CWA 15793 standard was introduced to 17 biotechnology-relatived microbiology laboratories. By focusing on continuous improvement and the Plan-Do-Check-Act (PDCA) cycle, this system will identify and monitor all aspects of laboratory biosafety and biosecurity.
Future Prospects
Taiwan CDC has participated in the Global Health Security Agenda (GHSA) for several years and realized the international concern of Biological Select Agents and Toxins (BSAT). In 2016, we applied JEE Tool (Joint External Evaluation Tool) published by WHO to assess the epidemic prevention system of our nation which includes reviewing the level of BSAT management. For the JEE Tool, the highest score for each indicator is score 5-sustaninable. Taiwan was scored 3-developed capacity in BSAT management. As a result, by 2018, Taiwan CDC has improved the insufficiency of BSAT management and has made a great progress on it. We believe by the time of 2020, we will reach a higher score on BSAT management on JEE tool, to prove the nation’s ability of managing BSAT can be as well as other advanced countries.
What our team did to comply with it
Not only did our team complied with the rules, we aimed to contribute to elevate the biosafety of lab and public health. In the section of design, we developed an AI system that is able to predict the spread of influenza.
To sum up, since our experiment involves in virus experiments, we paid special attention towards this issue to secure the safety for both the researchers and public.
Images 1-2. On the left, Roger explaining Marc how to deal with chemical containers, on the right, the containers which CGROM manage.
As an additional safety measure we use SYBR Safe instead of Eidium bromide. Its use is identical in all the protocols but without carcinogenic risks.
Access to the laboratory
As we are a multidisciplinary team formed by biotechnologists, engineers and artists, not everybody knows how to behaviour in a laboratory. That is why we did several seminars directed by Alejandro Vignoni and Yadira Boada to teach the basics of their laboratory. However, as a security measure, they decided that not everybody would be allowed to work in the lab alone, only biotechlogists and biomedical engineers could be there without surveillance due to their experience and knowledge adquiered during their degree.
Over the course of these seminars we were informed about the evacuation protocols, waste management and our rights and liabilities as UPV staff. These information can be seen in the information sheets they gave us and that you can download through these buttons:
Strains
Printeria uses E.coli and V.natriegens as chassis. Both are classified as risk 1 so the harm which can cause to people is minimal. The E.coli are from strains DB3.1, 10G and TOP10, for V.natriegens we use the strain ATCC 1404 from CECT (Colección Española de Cultivos Tipo). To do constructions we use biobricks and parts supplied by IDT prepared with the Golden Gate grammar.
Lab safety
Experiments regarding proteins of our iGEM project were conducted in the Biosafety Level-1 laboratories, whereas experiments associated with virus were conducted in the Biosafety Level-2 Laboratories. Both labs are under the regulation of Environmental Safety and Health Center of Chung Shan Medical University. The emergency exits and basic risk assessment are the first things to check every day prior to experiments.
Training
All members of the laboratory were well-trained by the mentors and had ability to conduct experiments in a safe environment. People who entered the laboratory should follow the established laboratory safety principles of our school. Master’s degree students of the lab and our instructor helped us to complete the safety training.
Special courses
Conference for Lab Safety, Microscope and Common Equipment Usage held by Department of Biomedical Sciences
By attending the conference, we learned several important issues regarding lab safety as listed below:
1. Safety precautions and operating notice of commonly used instruments:
The order to start the machine is of great importance. First, we should turn on the machine in the middle to warm it up for about 30-60 minutes. Then we turn the pump open. Note that if any water drop slides into the liquid, the pump will rise. Therefore, remember to drop the water droplets slowly from below.
Balance is very important! Balance the weight before putting the samples into the machine. Do not mess with the tools provided by a different company from the machine manufacturer. For example, if we want to centrifuge with 15 cc centrifuge tube, we use the centrifugal sleeve provided by the manufacturer instead of plugging the tube into any other centrifugal sleeve. Under high speed centrifugation, unbalance could cause great damage to the middle shaft of the machine (which is the heart of it), and would be very dangerous to operators since the samples could burst out from the machine in a very high speed. Don’t put any thing on the machine when it is operating as well.
The flow should be turned on for at least 20 mins before usage. The hood must be pulled down when the UV lights are on
Don’t put any PC materials into the machine or it could melt. Always remember to check if both the drainage knob and exhaust knob has been tightly closed before powering up the machine. Ensure that the door is fully sealed, however, don’t over-tighten the bolt as it may result in damage to the gasket. Before taking out the materials, turn the exhaust knobs loose. Gloves are necessary.
The volume of water column needs to be adjusted according to the black indicator line. The water will drain out if it exceeds the line .
Items must be placed in the middle of the scale. Always check the gradienter to see if the scale is standing at a horizontal platform .
2. School institutions that we can consult with when facing questions with the safety of experiments
˙ Always feel free to ask for help at the Environmental Safety and Health Center! If there is any problem regarding the instruments, please call the department for help, and the department will contact the company to handle it.
3. Other suggestions for improving the safety of laboratory operations:
If you’re uncertain about any step during the experiment, STOP! Never feel embarrassed to ask for help !!
MSDS
Material Safety Data Sheet, MSDS. Every chemical, reagents in the lab are printed and collected in this data folder. The folder is placed in a significant place near the door, which is easy to find and read in case of emergency.
Biohazard waste Management
All kinds of waste produced in the working area of the laboratory are considered biohazard waste, including every experiment and consumptive material. Those biohazard wastes would be collected into a specially labelled red plastic bag, and be sent to the Environmental Safety and Health Center of Chung Shan Medical University for further disposure process.
|
Safety Problem |
Implemented solution |
|---|---|
|
Sterilization of the PCB |
How do we clean the PCB (Printed Circuit Board) without damaging metalic and plastic components? Well, we decided to copy what flow cabins do. Based on this, we decided to use UV led lights to achieve this objective. This idea was given by Mojica as a part of our intergrated human. Furthermore we have programmed an especial reaction that use little droplets with HCl and ethanol to clean and ensure we have perfectly sterilized the PCB which is protected by a plastic film as well and this feature prevents acid from attacking it. |
|
Sterilization of the microfluidic tubes |
The reaction fluid is going to pass through microfluidic tubes and it will contaminate them. To reverse this situation we will use the same special program wich usses droplets with ethanol and HCl. |
|
Protection of the user |
We want users to see what is happening inside Printeria but also we want them to be protected if something wrong happens inside. That is why we thought that a metacrilate box that covered the machine would be a nice option. Besides, it is useful to pin up the elements which will be part of Printeria. |
|
Overheating of the PCB |
As the PCB has a cold and a hot zone, it is necessary to evacuate the heat generated. The hot zone can achieve more than 80º degrees and can be a potencial danger. A resistance is responsible of heating, and a peltier plate is responsible of chilling. Peltier effect is based in a semiconductor metal where you apply a potential difference. Then, you create a hot part and a cold one in that semiconductor. We use the cold part to extract the energy and stabilize the temperature via conduction of the PCB's hot zone. |
|
Ventilation |
The peltier plate is responsible of chilling the PCB but how do we evacuate all the energy generated? Well, the answer is simple, Printeria just needed ventilation holes and watercooling to do it. Watercooling is based in the higher termic capacity of fluids (water in this case) to absorb energy. Water is pumped through microfluidic tubes to reach the peltier and chill it. |
|
Electroporator |
This device is capable to reach high voltages, so if you require one of these in your design first of all, ensure no one can harm himself/herself. This is the reason why we designed our own electroporator in a dedicated and compacted printed circuit to isolate it from any exposure. It can reach until 1600V, which is not a joke. |
|
Tips |
Tips which go in the revolver wheel are disposable to ensure the purity of each reaction. |
Regulations
Science without ethics is cruel, and ethics without science is empty. Ethics become really important when a project is being thought. For this reason we wanted to pass the UPV (Polytechnic University of Valencia) ethics committee. Here you can download the document we sent them to approve and the positive veredict of the committee.
Ethics
Science without ethics is cruel, and ethics without science is empty. Ethics become really important when a project is being thought. For this reason we wanted to pass the UPV (Polytechnic University of Valencia) ethics committee. Here you can download the document we sent them to approve and the positive veredict of the committee.
References
1. Dual use: A report by iGEM team Bielefeld-CeBiTec 2015. [Germany]. Universität Bielefeld, . 30 pages.