Team:Grenoble-Alpes/Safety

NeuroDrop

Safety

Safe Project Design

Biological Safety

For biological device safety, the strain BTH101 is in group 1 and doesn't have any way to produce cAMP which make them very unlikely to grow outside optimal conditions.

Our transformed bacteria (BT101) will express fusion protein in the outer membrane. These proteins should not be pathogenic or have any environmental risk. Aptamer does not have a known risk for environment or health.The modified amino acid does not have a known risk for environment or health.Nano luciferase does not have a known risk for environment or health and to be sure, after the assay BTH101 will be eliminated by increasing the temperature up to 120°C in the device.

Device Safety

Regarding our device safety, we had to use other methods. Obviously it has to work on sector current. It needs autonomous power transformers which aren't as user friendly and safe as our power sources from the lab. First of all, all the metallic parts were linked to the ground. Then, there is an electromagnet on the front of our device. As soon as the device starts protocols and starts to aliment motors and the electro-wetting device, the electromagnet is powered and closes the door.

This way, the device is fully enclosed within its plastic box, and so there is no risks to touch any surface exposed to electrical risks.

Safe Laboratory Work

Safety Level : Our lab is a level 1. We only use E. coli strains, which is a Risk Group 1 organism.

Safety Training

The whole team had a safety training with the TIMC-IMAG lab supervisors. Many topics were discussed from waste treatment to safety equipments by way of emergency call.

The Biosafety rules in our lab are:

  • Tidy and clean your lab bench with disinfectant.
  • Wear your lab coat and if necessary gloves and glasses.
  • The lab coats have to be sterilized by autoclaving and regularly washed.
  • Don’t wear ring, jewelry, scarf or other fashion accessories.
  • Don’t eat, drink, put make up on or chew your pen in the lab.
  • Wash your hands before and after manipulations with soap and water.
  • Be careful with waste treatment. All biological waste have to be disinfected with bleach and discarded in the yellow biological bin.
  • Reusable equipments have to be disinfected with bleach before being washed and autoclaved.
  • Contaminated liquids have to be inactivated with bleach and discarded in the biological liquid waste can.
  • Don’t pipet with your mouth, or sample a powder with your hands.
  • All accidents, spills and activities with GMOs or human material have to be reported in the repertory.

Lab responsibility and supervisors

While we were using the lab, we were under the supervision of our advisors and the lab technician Sophie. She was there to help us with reagents, some experiments and material demands. Moreover, Sophie introduced us the laboratory, explained how machines work.
In addition, our supervisors, Mr. Pierre Cavailles, Audrey LeGouellec and Claire Durmort are teacher-researcher who accompanied us and gave us advices. These people were essential because of their experience and expertise, they allowed us to be in good and safe working conditions. Our supervisors are microbiologists, so they are familiar with the experimental procedures we use. In addition, they have been working for at least 5 years in the laboratory, so they have the reflexes and habits of work to optimize handling and safety.

Safety equipments

Each team member has a lab coat. Depending on our activities, we have to wear appropriate gloves (Nitrile, latex, heat-resistant) and we have glasses available. We avoid to wear skirts, shorts or sandals in the lab.

Lab equipments

We have to use many equipment in the lab as centrifuges, PCR machines, spectrophotometer...The first day we all had a training to learn how each equipment works. However if we have any doubt, we always could call Sophie or an advisor to obtain explanation before using anything.

Chemical reagents

We had to use many chemicals during our project. Before use, we read each material safety data sheets. Some reagents needed to be used with nitrile gloves and glasses.. The reagents that were dangerous (SGH08) were used in a chemical fume hood.

Waste treatment

In the lab we have three differents bins. They only are intended for solid garbage. The yellow one is for biological waste. Before discarding a biological waste, we had to disinfect it with bleach. The blue one is for cardboard. Then, the dark one is for the other kind of waste. We have to be careful not to discard our eppendorf tube or gloves in the dark one even if they are not in contact with biological waste.

Biosafety

Organisms

For our project we are using E. coli only. However we needed several different strains.
We used :

  • E. coli DH5-alpha, to transform parts contained on iGEM plates.
  • E. coli BL21, to visualize nanoluciferase expression.
  • E. coli BTH101 which doesn't express Adenylate cyclase for our final project.
All of them are Group 1 Bacteria. They are not dangerous for an adult healthy human.

Plasmids

For our project we used pSB1C3,pSB1C00 and PSB1A2 vectors. We used a commercial vector named pUT18 with which we built our final plasmid.

Antibiotics

For our project, we used four antibiotics : ampicillin, kanamycin,streptomycin and chloramphenicol.

Engineer Lab Safety

Concerning our safety in the engineering lab, things were very straight. As we didn’t use any chemicals and were helped by our fablab to manufacture pieces, our lab was very safe. Indeed, there was only one type of risk in the lab: electrical hazard.

But our luck was to work in a electronic school. So instead of having to work with hacked alimentations, all of our experiments were done on regulated alimentation. These devices are power sources that can be limited on current or voltage. By using this, even if our manipulations were done under high voltage, we weren’t exposed to any shocking hazard.

Obviously, all the time they were running, the power sources were kept under 40mA max intensity, and they were shutted off each time we had to put our hands on the device. In emergency case, the whole lab was equipped with an emergency circuit opener to shut down all the electrical alimentation in the lab.