Team:Lambert GA/Safety

TEAM LAB 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 get autoclaved
    • Check all equipment for good working order, no chips, torn cords, 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 cabinets
    • 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

Recognizing the current challenges around the world concerning the diagnosis of diseases caused by helminth worm infections, Lambert iGEM is creating a species-specific helminth detection method by developing a toehold switch biosensor, specifically for Caenorhabditis elegans. The C. elegans toehold switch can serve as a model design for more toehold switches that can be applied to other helminth species to detect the pathogenic organisms.

Caenorhabditis elegans is not generally considered capable of infecting vertebrates, and therefore the risk of accidental human infection is negligible. The risk of exposure and infection to the workers in the lab is very low, and good standard laboratory practices, safety equipment, and disposal protocols will prevent any accidental exposure of the human skin to Caenorhabditis elegans.

BL21 and DH5-alpha E. coli is nonpathogenic and was developed for laboratory cloning use. The potential health and environmental hazards associated with BL21 DH5-alpha E. coli hazards are highly limited and can be handled in Biosafety level 1 laboratories.

Hardware/Software

3D Printer

The main safety hazards with our 3D printer involve temperature fluctuation. The nozzle is able to heat up to 220℃. Because of this, leaving the printer unattended to can be dangerous due to the 3D printer’s high temperatures, which could potentially cause a fire. However, our printer possesses mechanisms that prevent this from happening, so the chance of fire is extremely low. If a fire does occur, it can burn parts of the printer and damage anything around it. The printer uses software called Crash Detection and Heat Overload to monitor the overall safety of the printer. The printer will always be monitored by a PI to prevent any unwanted accidents.

MicroMesh

MicroMesh does not pose any hazard to the user after it is built, but precautions have to be taken while making it. MicroMesh is made from larger mesh sheets, which poses a sharpness hazard for the mesh. When the mesh is cut, the edges could be sharp and it can poke fingers, especially with the largest strainer on top. For our purposes, we had an extra precaution by hot gluing the edges so they would not be sharp before gluing it to the top of the funnel piece. Also, fecal matter is a hazardous material, so during our time at Emory when we worked on testing if MicroMesh worked with fecal samples. We tested inside fume hoods to ensure that we did not contaminate the lab and all waste was disposed in biohazard waste.

MicroMesh is very safe to use once made. But, while working with MicroMesh, it is important to be careful with any sample that is being dealt with. The samples are most likely to be hazardous since there is a high possibility that they contain helminths or helminths eggs in them. Additionally, if the sample being dealt with is fecal matter, then additional precautions would have to be taken because fecal matter is inherently hazardous.

In order to clean MicroMesh one must use a bleach solution to rid the filter any remaining biohazard components. When using the bleach solution make sure that it is a 1:10 solution and that the filter soaks for over an hour and an overnight soak ensures maximum efficiency. We reccomend a solution with 30 ml of bleach and 270ml of water to soak MicroMesh filters in. After this is done, MicroMesh may be reused for filtering. But to ensure the filter is efficiently clean, run water through the filter and remove any excess solids. If the flow of water is constant and it is not being blocked by any components, MicroMesh remains safe to use. In our biosafety level one lab, we did not test fecal samples, however, during our time at the Barr and Ryan labs at Emory University, we utilized this precaution to ensure MicroMesh was safe to use again.

OpenCellX System

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 human skin, the voltage is not high enough to cause any real damage to a person.

Although OpenCell does spin at extremely fast speeds, all of the gearing is contained and thus eliminates any chance for accidental contact with a user’s fingers. Therefore, if OpenCell is used correctly with the lid, any risk to the user will be mitigated. Using our protocol, OpenCell will allow the user to run a full DNA extraction with very little risk to the user.

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 possible, for the consumer to print both OpenCell and OpenCell Pro in ABS (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 of electrocution.

The OpenCell Pro device utilizes a much stronger current and higher voltages than those of the frugal Opencell 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 connects have been properly soldered and wrapped with heat shrink to prevent short-circuits or disconnections, and all electronics are sealed in a separate section of the machine. In the case of malfunction, 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 that the device is made from, a layer of white lithium grease is applied to reduce friction and create an efficient system. Bearings, M3 screws, and denser designs are also included to create a safe machine that can be used in both labs and in school.

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.

Ethical Cultural Exchange Practices

Throughout the entirety of the Labyrinth project, consideration was given to team practices that could impact invested stakeholders. In particular, the Lambert iGEM team invested in academic interchange which fostered information exchange between Dominican high school students and American high school students. Specifically, Lambert iGEM students focused on lessons centered around frugal microscopes (Foldscopes) and the relationship between personal hygiene and disease transmission, while students from Santa Maria del Batay and Marcatcho High School taught us about homeopathic remedies for common ailments, including parasitic infections, menstrual problems, and diabetes.

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 [2].

REFERENCES

[1] AnimalResearch.info. (2014). C. elegans (nematode worm). Retrieved from http://www.animalresearch.info/en/designing-research/research-animals/c-elegans-nematode-worm/.

[2] 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

[3] 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

[4] Rochester Institute of Technology. (2019). 3-D Printer Safety. Retrieved from https://www.rit.edu/fa/grms/ehs/content/3-d-printer-safety.