Team:Sydney Australia/Safety

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Safety

Being safe in the lab is of utmost importance to us in the Magi.Coli team. We work in a PC2 space so whilst we are working with a relatively safe organism (cannot propagate outside of lab conditions and has a very low-likelihood of infecting immune-competent individuals) we still have to be careful to not contaminate ourselves with the organisms our colleagues are working on!

Before even stepping foot into the lab we needed to complete an online training course on PC2-level safety procedures to learn about topics including biological/chemical waste management, hazard identification and reduction, safe use of equipment, and emergency evacuation procedures. We then all underwent an in-person induction to confirm our knowledge of safe-work procedures, appropriate use of PPE, and safe use of the specific equipment available in our lab

The door to the shared lab and the PC2 certification and lab safety guidelines

Every day we go into the lab to do our work we need to be aware of the risks we undertake and how to prevent them using the following safe-work practices:

  • Personal Protective Equipment (PPE): As soon as we enter the lab we don our own lab coat, safety glasses, and gloves. Gloves must be worn while touching anything in the lab not in a designated ‘green zone’.

  • Green zones: Green zones in the lab are specific sterilised areas for us to keep things we do not want to get contaminated such as our lab book. Gloves are not allowed to be worn when interacting with objects in green-zones to keep them contaminant free.

  • Ethanol sterilising surfaces: Before and after lab work is conducted on an open bench, surfaces must be sterilised with ethanol or F10SC.

  • Using Bunsen burners to create sterile environments: When conducting microbial work on an open bench, a Bunsen burner is turned on to create an updraft, keeping the area sterile and preventing cross-contamination.

We do a lot of potentially hazardous things every day, here’s a list of some of our most common risks and how we mitigate them:


Risk: Electric shock from electrophoresis apparatus due to exposed/damage wires or electrodes.


Safety behaviour: check electrophoresis apparatus for signs of damage before using the equipment. If the equipment looks damaged alert the lab manager and do not use the equipment. Do not open the electrophoresis apparatus whilst it is running. Training on safe use of the equipment.



Risk: Burns from hot agarose.


Safety Behavior: Only handle hot agarose bottles with heat-proof gloves.



Risk: Cancer/burns risk from UV light from transilluminator.


Safety Behavior: Use an gel-imager that is fully sealed and does not turn on until the doors are completely closed. Do not use the imager if it is showing any signs of damage as it could result in malfunction of this safety feature.


Risk: Possibility of release of GMOs to the environment due to incorrect use of cabinet.


Safety behaviour: All students have been trained on appropriate use of the biosafety cabinet including how to prevent obstructing airflow, how to know when the sash is placed correctly, and more.


Risk: Possibility of fire in the biosafety cabinet due to inappropriate use.


Safety Behavior: All students have been trained on appropriate use of the biosafety cabinet including not putting flammable materials in the cabinet and not utilising a Bunsen burner inside the cabinet.


Risk: Possibility of burns to eyes or skin from UV light in cabinet.


Safety Behavior: All cabinets are calibrated to not allow UV light to turn on unless the cabinet is fully closed, the glass of the cabinet blocks UV light from exiting the cabinet. Students are trained to not try to place body parts inside the cabinet while the UV light is on or to attempt to turn on UV light while the cabinet is open in case of malfunction of this feature.


Risk: Fire hazard due to use of ethanol to sterilise the cabinet.


Safety Behavior: All students are instructed to sterilise the cabinet using F10-SC or to spray ethanol on a paper towel outside the cabinet before wiping down to prevent excess ethanol inside the cabinet.


Risk: Bunsen flame can cause burns.


Safety behaviour: Students are instructed to keep long hair tied back while in the lab and to not place any body parts near the flame.


Risk: Bunsen flame can start fires if flammable materials are nearby.


Safety Behavior: Students are instructed to only turn on Bunsen burners in spaces clear of flammable materials during their safety induction.


Risk: Risk: Rotor imbalance can cause catastrophic failure of centrifuge (explosion), injuring user.


Safety behaviour: Students are trained in appropriate loading of the centrifuge during their lab induction.


Risk: Rotor can fail due to extensive wear and tear, causing catastrophic failure of centrifuge, injuring user.


Safety Behavior: Students are instructed to look for obvious signs of wear and tear before loading their samples into the centrifuge. Machines with wear and tear will be tagged out and lab managers notified.


Risk: Centrifuge tubes can break in rotor due to excessive wear and tear or inappropriate use, causing an imbalance of rotor, catastrophic failure of centrifuge, injuring user.


Safety Behavior: Students are instructed to always use appropriate tubes for their sample and to not use broken or cracked tubes inside the centrifuge to prevent breakage.


Risk: Contact/Absorption - corrosive liquids have high potential to cause serious burns and irritation to skin and eyes.


Safety behaviour: Students wear gloves and safety glasses at all times within the lab to prevent skin contact with corrosive materials. Students are instructed to change gloves and dispose of them in the chemical waste bin after handling corrosive substances.


Risk: Inhalation - corrosive gases/vapour can damage the lining of the lungs.


Safety Behavior: Students are instructed to use any chemicals that have a risk of inhalation inside a fume hood to prevent inhalation.


Risk: Severe irritation (nausea, vomiting and diarrhoea) and permanent damage to the gastrointestinal tract.


Safety Behavior: Students are trained not to ingest any substances inside the lab.


Risk: May react with other substances and generate/evolve heat.


Safety Behavior: Students are instructed to read appropriate safety documents before combining corrosive substances and to not mix substances without a well-researched protocol.


Risk: Eye damage if corrosive chaotropic salts contact eyes.


Safety behaviour: Students wear safety glasses at all times within the lab


Risk: Poisoning if chaotropic salts ingested.


Safety Behavior: Students are trained not to ingest any substances inside the lab


Risk: Skin damage if chaotropic salts contact bare skin


Safety Behavior: Students wear gloves and lab coats at all times inside the lab to minimise exposed skin.


Risk: Environmental damage if chaotropic salts disposed of incorrectly.


Safety Behavior: Students are instructed as to how to appropriately dispose of both their biological and chemical waste into appropriate biohazard and chemical waste bins respectively


Risk: Environmental damage if recombinant DNA is disposed of incorrectly.


Safety Behavior: Students are instructed as to how to appropriately dispose of both their biological and chemical waste.


Risk: Injury due to incorrect use of centrifuge.


Safety Behavior: Students are trained in appropriate use of the centrifuge during their lab induction.


Risk: Fire or electrocution due to centrifuge malfunction.


Safety Behavior: Students are instructed not to use the centrifuge if it shows any signs of wear and tear. Malfunctioning centrifuges are tagged out and reported to the lab manager.


Risk: Harmful vapours (toxic, corrosive, stench) may escape from the hood if it is not set up correctly.


Safety behaviour: Students are trained in how to safety set up a fume hood during their induction.


Risk: Large objects can disrupt the airflow within the fume hood.


Safety Behavior: Students are instructed not to place large objects inside the fume hood.


Risk: Use of a Bunsen burner or other ignition source in hood can lead to a fire.


Safety Behavior: Students are instructed not to use Bunsen burners or other sources of flame within the fume hood.


Risk: Use of substances such as acrylamide, SDS (sodium dodecyl sulphate), TEMED, APS, Mercaptoethanol, Methanol, and Acetic Acid - potential for inhalation, skin/eye contact, ingestion. Some of these substances are also highly flammable.


Safety behaviour: Students wear lab coats, safety glasses, and gloves at all times within the lab to prevent contact of exposed skin or eyes with these substances. Students are taught not to consume any substances within the lab. All gels are bought pre-made to prevent students from handling any of the substances themselves. SDS-PAGE gels are prepared away from flame.


Risk: Potential for serious electrical shock or electrocution due to leaking chamber, faulty or corroded electrode cables, or faulty power supply.


Safety Behavior: Check electrophoresis apparatus for signs of damage before using the equipment. If the equipment looks damaged alert the lab manager and do not use the equipment. Do not open the electrophoresis apparatus whilst it is running. Training on safe use of the equipment.


Risk: Proteins samples. May be recombinant in nature and/or may have other hazardous properties (e.g. toxins).


Safety Behavior: Students do not touch the protein samples directly. All protein samples are disposed of in appropriate waste containers.


Risk: Chipped glass plates. Potential for causing cuts to skin. May also result in leaking of un-polymerised acrylamide solution.


Safety Behavior: Students are instructed to look for any chips or cracks in the glass plates before commencing an SDS-PAGE.


Risk: Loading samples via needle syringe. Potential for needle stick injury.


Safety Behavior: When using needles students are instructed to dispose of used needles in sharps bins and to not attempt to re-sheath the needle.


Our unique risks

Our project involves the synthesis of a restricted substance, psilocybin. The first step in making sure we were being safe and complying with our state and university regulations was to notify them and get appropriate licensing for production and possession of psilocybin and its intermediates, norbaeocystin and baeocystin. Sam Bannister from the Lambert Initiative helped our supervisor Nick Coleman with the paperwork required by the NSW Government. A copy of abridged versions of the approval papers from the NSW Government and from our university.

Approval documents from the NSW Government and USYD Instituitional Biosafety Commitee

Additionally, it has been incredibly important to us that we're all aware that working with this restricted substance comes with its own unique set of risks:

    • Risk: Accidental ingestion or overdose
    • Safety behaviour: In terms of acute lethal toxicity, psilocybin is a relatively safe drug. An average adult would need to ingest about 20 grams for a lethal dose, which equates to 17 kg of bacterial cell pellets, which equates to approximately 10,000 litres of culture. Even for a psychoactive dose an individual would need to consume about 20 mg of psilocybin, which equates to one litre of raw culture. As students are instructed not to consume any substances produced or contained within the lab it is extremely unlikely that this risk would eventuate, especially as we only plan on making at maximum one litre of culture at a time.

    • Risk: Legal risks if our organism was not contained under NSW law - i.e. individuals without appropriate licensing could either inadvertently or deliberately posess our psilocybin containing E.coli.
    • Safety behaviour: our organism is contained within our lab using biosafety procedures. Students are not allowed to take culture out of the lab, and hands are washed and garments removed to prevent any accidental transfer of our organism to the outside world.

    • Risk: Security and safety risks such as theft or illicit use.
    • Safety behaviour: Our lab floor is key-card restricted to only researchers who work within the building, and the lab itself is further restricted to only individuals working on our floor. All entries and exits are logged so any theft would be easily caught and the culprits identified. Students are also instructed not to bring any visitors into the lab space, especially when we are working with restricted substances.

We have also performed a literature review on the risk our final product - psilocybin - would hold within the community were it to be released as a therapeutic. Please see our page: Human Practices - Safety of Psilocybin for more information.