Safety

Team Safety

All team members involved in lab procedures were required to adhere to certain lab guidelines as outlined by our advisors. The lab safety rules include but are not limited to:

• 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

Public Safety

Lambert iGEM took certain measures to ensure that our project did not pose a threat to public safety. All E. coli strains handled were non-pathogenic and are unlikely to survive in the wild, even if released. All biomaterials were disposed of properly and all relevant lab equipment were autoclaved after use. Team members were not allowed outside of the lab while wearing gloves and refrained from touching doorknobs or any surfaces that would come in contact with the public. The team also cleaned all lab surfaces after use to ensure that others did not come in contact with our biomaterials.

Environmental Safety

To ensure that the project did not propose a hazard to the environment, Lambert used only transformation strains that were purchased through New England Biolabs or from the CSCG Keio Collection. The project aim is to further characterize the degradation strength of two degradation tags, DAS and LAA, and as such, the cell lines will never leave the lab, and they will not pose a threat to the environment because they are non pathogenic and contained in a laboratory environment.

Decontamination and Sterilization Protocol

Introduction

Decontamination has a vital role in ensuring safety and health by preventing infection and disease. In any healthcare or research environment, sterilization has become a necessary step to minimize risk of contaminated equipment or patients. The following procedures establish a basis of accountability assuring us with guidelines that would appropriately decontaminate equipment and supplies before use to effectively allow the lab protocols to be accomplished accurately. The Chrome-Q Light Chamber is a 3D-printed imaging measurement system used to quantify results of nutrient deficiency. To use the Chrome-Q, first a sample must be placed on the base under the center of the dome. Then, using the app, a phone can take a picture of the sample through the eyehole, and calculate degradation of protein by analyzing the RGB values in the pixels of the samples. By following the procedures below, the Chrome-Q will become cleaned and disinfected to provide a sterile environment for the colored samples to assure accurate readings.

Definitions

Cleaning is the removal of organic and inorganic material from objects and surfaces. Cleaning is accomplished by utilizing detergents or enzymatic products. Since organic and inorganic materials that remain on the surface of instruments interfere with the effectiveness of these processes, thorough cleaning is required before disinfection and sterilization.
Decontamination involves the physical or chemical means to remove, inactivate, or destroy microorganisms on a surface or item is rendered safe for handling, use, or disposal. The selection and use of cleaning equipment, chemicals, and exposure times suggested by the device manufacturer should generally be followed to prevent damage to the items.
Disinfection is a process that reduces the number of microorganisms (with the exception of bacterial spores) on inanimate objects. This is done most often by use of an approved hospital detergent/disinfectant or chemical sterilant.
Antisepsis inhibits the growth of microorganisms on living tissue (e.g., skin preparation before vascular line insertion or other invasive procedure). Alcohol, chlorhexidine gluconate and lodophors, i.e., betadine are the most frequently used solutions for antisepsis. Germicidal chemicals used for antisepsis are not generally adequate for decontaminating environmental surfaces.
Sterilization is the complete destruction of all microbial life. It is accomplished by either a physical or chemical process, such as steam under pressure, dry heat, ETO gas and liquid chemicals. All items that enter sterile tissue or the vascular system must be sterile, i.e., implants, scalpels, needles, surgical instruments, etc.

Procedures

1. Pre-cleaning: Contaminated items should be wiped with Clorox Wipes to reduce the number of microbes and to keep the objects moist prior to cleaning.
2. Use a disinfectant: Wipe the Chrome-Q off with a clean cloth soaked in iodine, povidone-iodine (Betadine) or Ethanol alcohol.

Severe Surface Decontamination

1. Prepare a household bleach solution (1 ½ cups of bleach with 1 gallon of water)
2. Place the solution in a spray dispenser.
3. Thoroughly douse surfaces that have heavy deposits of contaminants allow to stand for 3 minutes.
4. Wipe the contaminants and allow to stand for 3 minutes.
5. Wipe the contaminants from the surface with a paper towel.
6. Discard the towel.
7. Douse the surface again but utilize a weaker bleach solution and wipe off the residual contamination with a paper towel.
8. Discard the towel and allow surface to dry.

Sterilization Protocol for Chrome-Q

1. Clean: mechanically clean internal and external surfaces, including brushing internal channels and flushing each internal channel with water and a detergent or enzymatic cleaners.
2. Disinfect: immerse Chrome-Q in a disinfectant (or chemical sterilant).
3. Rinse: rinse all channels with sterile water or filtered water.
4. Dry: rinse the Chrome-Q with alcohol and dry with forced air after disinfection and before storage.
5. Store: store the Chrome-Q in a way that will prevent recontamination and will promote drying (e.g., hung vertically)

Gaurav Byagathvalli practicing proper safety in the lab.