Team:Lambert GA/Safety
TEAM SAFETY
- Wash in, wash out
- Protect eyes, mucous membranes, open cuts, and wounds from contact with biohazard material
- Do not eat or drink when in the lab area
- Always use gloves and splash-proof goggles
- Tie back loose hair
- Disinfect all surfaces with 70% ethanol prior to working
- Disinfect all disposable tips, glassware, tubes by soaking in 10% bleach solution for 20 minutes and then disposing in normal waste
- Dispose of growth plates by disposing into a biohazard container which gets autoclaved
- Check all equipment for good working order, no chips, torn chords, cracks. Report any issues to an instructor immediately
- When pipetting, don’t touch tip to side of container
- Don’t lay caps of tubes upside down. Use masking tape to hold to bottom of cabinet
- Clean work area with 70% ethanol after working
- Clean up all glassware and labware before leaving lab
- Place all backpacks and stools to the side of the lab to keep walkways clear
- Always know the correct procedure for disposal of lab materials
PROJECT SAFETY
OVERVIEW
As the third largest food desert nationwide, Atlanta, GA faces a high number of diet-related health issues. In order to combat this local issue, Lambert iGEM aims to create a low-cost aquaponics system utilizing biosensors to detect concentrations of nutrients essential to plant growth such as phosphate and nitrate. The biosensors are designed to express different levels of GFP in response to nutrient volume presented in the aquaponics system. Specifically, Lambert iGEM will characterize a phosphate biosensor created by NUS Singapore iGEM 2017, part BBa_K2447000, which takes advantage of the natural E.coli Pho regulon. For our nitrate/nitrite biosensor, the team will characterize part BBa_K1682018, previously created by HKUST-Rice iGEM 2015, that utilizes the traditional Nar operon proteins. The E. coli containing the biosensors will be isolated in a compartment to avoid contamination of the rest of the aquaponics system. BL21 and DH5-alpha E. coli is nonpathogenic and was developed for laboratory cloning use. The potential health and environmental hazards associated with BL21 and DH5-alpha E. coli are highly limited and can be handled in Biosafety level 1 laboratories.
HARDWARE/SOFTWARE
3D PRINTER
The biosensor cells responsive to nutrients are planned to be used within a 3D-printed chamber for an aquaponics system. The main safety hazard with our 3D printer is the temperature fluctuations. The nozzle heats up to 220 degrees Celsius. In addition, leaving the device unattended can be dangerous due to the 3D printer’s high temperatures that could cause a potential fire. However, our printer is advanced and possesses mechanisms to prevent this from happening, so the chance of this occurring is extremely rare, but if it occurs, it can cause serious damage and end up burning parts of the printer. The printer uses software called Crash Detection and Heat Overload to monitor the overall safety. The printer will always be monitored by a PI to prevent any accidents.
OPENCELLX
OpenCell utilizes household 9V batteries to run the 12V DC motor. Because all of the electronics on OpenCell are contained, the safety risk is minimal and due to the resistance of skin, the voltage is not high enough to cause any real damage. Although OpenCell does spin at extremely fast speeds, all of the gearing is contained and thus eliminates any chance for accidental contact with the user’s fingers. Additionally, during all testing, a shatter proof lid/enclosure is used in case of catastrophic failure. Using proper safety protocols, properly using the lid, and the correct batteries, there is very little risk of injury when using this device. Furthermore, the 3D printed parts are stable enough to run for extended periods of time so the risk of failure mid-spin is minimal and very little long-term maintenance is required. Our design utilizes PLA or Polylactic Acid which is a common, non-toxic 3D-printing compound. We recommend, if it is possible, for the consumer to print both OpenCell and OpenCell Pro in ABS or Acrylonitrile Butadiene Styrene due to its increased strength and long-term durability. Because OpenCell is our frugal design, we have spared some optional safety mechanisms to increase the portability, frugality, and in-field practicality of our device. However, OpenCell Pro has numerous safety features detailed below that ensure the user will never come into contact with the wired electronics, significantly reducing the risk for any type of electrocution. The OpenCell Pro device utilizes much stronger current, higher voltages, and employs an array of electronic components to function. As a result, special precautions have been taken to ensure that the device maintains acceptable safety standards during operation. On the electronics side, all connections have been properly soldered and wrapped with insulation material to prevent short-circuiting or disconnecting, and all electronics are sealed in a separate section of the machine that is not electrically conductive. In case of any failure, a master power switch is included to shut off all power to the device. Mechanically, the OpenCell Pro operates at much higher speeds than the base model, and to prevent the degradation of the thermoplastics and reduce the risk of component failure, a layer of white lithium grease is applied to reduce friction and reduce wear on any moving surfaces. Bearings, M3 screws, and stronger design elements are also included to create a lab, and school, safe machine.
ELECTRONIC COMPONENTS
For this project, there is a significant amount of electronics work that will be done and we have taken all of the proper safety precautions. Before any work with arduino, raspberry pi, or related sensors and power supplies, team members ensure that they ground themselves with a large metal object. During any electronics work, team members use safety glasses. If working with motors or when soldering team members wear electrically and thermally insulated gloves and work in a seperate part of the room to reduce any risk of injury to other team members. Special precautions are taken when using any power supplies and AC adapters including inspecting power supplies for damage before use, ensuring that proper voltages and currents are provided, and all liquids, food items, or other conductive materials are kept away.
HUMAN PRACTICES
Participants in all surveys provided signed consent for the release of their responses from themselves or a legal guardian. Participants in all events hosted by Lambert iGEM provided consent for photo and video release.
DISCARDING CELLS IN THE FIELD
A solution of 1% sodium hypochlorite and 70% ethanol will kill biosensor cells. Results show that a 1% sodium hypochlorite solution sprayed on the surface and let sit for five minutes will effectively remove all DNA, saliva, blood, semen, and skin cells from any smooth or pitted surface when wiped down with 70% ethanol afterward. However, sodium hypochlorite solution followed by ethanol can produce amounts of gaseous chlorine above recommended exposure levels. As a result, 1% sodium hypochlorite followed by distilled water was tested and proven to be effective as well (Kaye et al., 2015).
REFERENCES
Kaye N. Ballantyne, Renato Salemi, Fabio Guarino, James R. Pearson, Dale Garlepp, Stephen Fowler & Roland A.H. van Oorschot (2015) DNA contamination minimisation – finding an effective cleaning method, Australian Journal of Forensic Sciences, 47:4, 428-439, DOI: 10.1080/00450618.2015.1004195
National Institute of Health, Federal Select Agent Program. (2014). Biosafety and biosecurity in the United States. Retrieved from https://www.nih.gov/sites/default/files/research-training/usg-safety-factsheet-2014.pdf
Rochester Institute of Technology. (2019). 3-D Printer Safety. Retrieved from https://www.rit.edu/fa/grms/ehs/content/3-d-printer-safety.