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Bacterial Dyes - Bioshades

Learning paths

Bacterial Dyes, Working with microorganisms for pigment extraction and finding alternatives for chemical dyeing.

Path Target Duration Skills
PathX 18-25, ++ half a day Learning how to dye with bacteria

Introduction and context

One of the most emerging developments that is currently growing rapidly in the field of industrial design, fashion and art is the creation of biobased materials, bio-fabricated in laboratories from microorganisms, bacteria and biopolymers. The industry is already incubating solutions to create new products that propose alternatives to materials derived from petroleum or animals, with examples from small and large companies such as Deserto, Piñatex, Bolt Threads, Gelatex.

The ¨Bacterial Dyes¨ workshop is a demonstration and hands-on activity to create awareness on the origin of colors, and how natural dyes were replaced with chemical dyes in the past century. In this case we can explore the possibilities of microorganisms producing pigments, through a focused hands-on activity on some of the stages of the entire process.

This learning path is about Bacterial Dyes. It raises awareness around the origin of color and how to work with natural dyes.

The activity is designed for a moderate number of participants, for being a hands-on activity each participant will get the opportunity to exercise in different stages of the dye process under direct supervision. The ideal number is 8 to 12 people.

Learning Outcomes
- How to work in a DIY sterile environment - How to safely work with bacteria at Biosafety Level 1 (BSL-1)

Target audience and context of use
Creatives and professionals, such as producers, designers, architects, biologists, SMEs, students of creative fields of knowledge, and companies with interest in sustainability and alternative innovative biobased materials.

You can run the activity in different places like a university, a lab, a cultural center, a museum, etc. Set up a collaboration between your lab, material centres and a museum.

Part 1: BACTERIA PREPARATION

Preparation and materials

Equipment, Tools and Materials

  • 4 tables, chairs (one table central display, three for students)
  • Video recording and photo shoot set up for distance live or remote observation, if necessary.
  • Safety equipment: 95% Ethanol,Kitchen paper, Disposable gloves, Lab Coats, Goggles, Bleach, Oven glove
  • Autoclave bag
  • Glass Petri dishes
  • Inoculation loop, ideally steel.
  • Pipette
  • Pressure Cooker
  • Bunsen Burner (or camping gas stove) + Lighter
  • Incubator
  • 250 ml Reagent Bottles with blue lid
  • Permanent marker
  • Parafilm
  • Bacteria : Janthinobacterium Lividum
  • For 4 sterilized petris of 100 mm diameter :
  • 200 ml distilled water
  • 4 g Nutrient Agar
  • 0.5 ml Glycerine
  • Note: scale the amounts according to the total amount of petris to prepare.

Step by Step Instructions

STEP BY STEP overview

  • Dissolve the nutrient Agar in 200 ml of distilled water
  • Add 0.5 ml Glycerine with a pipette
  • Firmly close the Reagent bottle and shake until dissolved
  • Make 2 or 3 bottles depending on the amount of participants, calculate one bottle for every 3 or 4 participants.
  • Add approximately 1 litre of water in the pressure cooker
  • Place autoclave bag into the pressure cooker
  • Place the bottles vertically with the lid half unscrewed and the empty petri dishes closed into the autoclave bag.
  • Close and pressure cook for 30 minutes.
  • Release the steam carefully away from the face before opening
  • Clean the table with alcohol and paper
  • Pour alcohol in a circle
  • Place the Bunsen burner in the center and turn it on
  • Place the sterilized petri dishes around the burner without opening
  • Unscrew the bottle’s lid close to the burner and sterilise its edge placing it directly over the stove’s flame.
  • Quickly open the petri dish and pour 4-5mm height of the medium covering all the surface of the petri dish.
  • Let it rest until firm/cooled down
  • Use the same setup as step 3
  • Sterilize your hands (wearing gloves) with alcohol
  • Place the petri containing the bacteria near to the burner
  • Sterilize the inoculation loop by passing it through the flame (skip this step in case of single use plastic loops)
  • Open the petri and place the inoculation loop next to the bacteria to cool it down
  • Scrape gently the bacteria from one petri and transfer them to the other
  • Label each petri dish by writing down medium, bacteria name, date and person
  • Tip: Open the petri dishes towards the flame / sterile side
  • Seal the petri dishes with the parafilm
  • Place upside down (agar up) in the incubator at 24°C for 48 - 72 hrs.
  • Set up your working Area following step 3 using Personal Protective Equipment (PPE)
  • Pour 96% ethanol into the petri and extract the pigment into a jar.
  • To kill the bacteria, place the petris in an autoclave bag and pressure cook following step 2.
  • Close and dispose of the autoclave bag.
  • Sterilize your working area and wash your hands with alcohol.

Part 2: TEXTILE DYEING

Preparation and materials

Equipment, Tools and Materials

  • Needle and thread
  • Rubber bands
  • Textile (Hemp, Silk, Viscose)
  • For 4 petris of 100 mm diameter250 ml distilled water
  • 6.75 g LB Broth
  • 1 ml Glycerine
    Note: scale the amounts according to the total amount of petris to prepare.

Step by Step Instructions

STEP BY STEP overview

  • Dissolve 6.75 gr of LB Broth in 250 ml distilled water
  • Add 1 ml Glycerine with a pipette
  • Firmly close the bottle and shake until dissolved
  • Rinse and dry the textile beforehand
  • Fold & stitch the textile using tie dye, Shibori techniques
  • Place them inside the petri dish making sure that it closes
  • Add ±1 litre of water in the pressure cooker.
  • Place the autoclave bag into the pressure cooker
  • Place the bottle vertically with lid half unscrewed and the petri dishes with the textiles closed into the autoclave bag.
  • Close and pressure cook for 30 min.
  • Release the steam before opening.
  • Clean the table with alcohol and paper.
  • Pour alcohol in a circle.
  • Place the Bunsen burner in the centre and turn it on.
  • Place the petris around the burner without opening.
  • Unscrew the bottle’s lid close to the burner and sterilise its edge in the fire.
  • Quickly open the petri dish & pour the medium on the textile.
  • Let it rest until firm/cooled down.
  • Use the same setup as step 4.
  • Sterilize your hands (wearing gloves) with alcohol.
  • Place the petri with the bacteria close to the burner.
  • Sterilize the inoculation loop by passing it through the flame (skip this step in case of single use plastic loops).
  • Open the petri and place the inoculation loop next to the bacteria to cool it down.
  • Scrape gently the bacteria from one petri and transfer them to the textile. (without breaking the gelatin)
  • Label each petri dish by writing medium, bacteria name, date and person.
  • Tip: Open the petri dishes towards the flame/ sterile side
  • Seal the petri with the parafilm
  • Place the petri in the incubator at 24 C° for 48-72 hrs
  • To kill the bacteria place the petris open in an autoclave bag and pressure cook following the step 2
  • Take the textile with the tweezers out of autoclave bag and rinse
  • Sterilize the petris, your working area and wash your hands with alcohol
  • Close and dispose the autoclave bag

Safety rules & Tips

  • Wash your hands, then disinfect them with alcohol-based hand sanitizer before and after the process.
  • Use personal protective equipment (PPE): Gloves, Goggles & Lab. Coat.
  • Sterilise your working area before, throughout and after the process.
  • Work only with Biosafety Level 1 (BSL-1)
  • Don't touch your face or pull your hair back once the activity has started.

Note : The instructions are for reusable glass petri dishes that need to be pressure cooked and later on sterilized with bleach or alcohol. In the case of disposable plastic petri dishes place the textiles in a separate autoclave bag.

With the dyed fabric, depending on the size and proportions, you can make a scarf or a bandana, the borders of the fabric need to be sewn.

Flyer for participants

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Tips to facilitate the activity in context (to-do / not do)

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Estimated cost

We recommend purchasing tools and materials locally, the cost is approximate and the list serves as reference. You might plan some costs for getting the materials and tools but also some fees linked to your collaboration with the place you run the workshop.

Item Quantity Cost (€)
6 glass Petri Dishes 100 mm diameter 1u 70
20 Plastic Petri Dishes 100 mm diameter 1u 60
Distilled water 1l 1
Nutrient Agar 100g 28
LB Broth 100g 34
Glycerine 1l 13
Autoclave bag 100 47
6 glass Petri Dishes 100 mm diameter 1u 70
20 Plastic Petri Dishes 100 mm diameter 1u 60
Inoculation loop, ideally steel 10u 30
Pipette
Pressure Cooker 1u 50
Bunsen Burner (or camping gas stove) 1u 30
Incubator
250 ml Reagent Bottles with blue lid 2u 10
Permanent marker 1u 2
Parafilm 38m 32
Textiles (Hemp, Silk, Viscose)
Needle and thread sewing package 1u 7
Rubber bands
95% Ethanol 1u 5
Kitchen paper 1u 2
Pack of Disposable Gloves 1u 4
Lab Coats 1u 25
Goggles 1u 3
Bleach 5l 12
Oven glove

References

Links to photos and recorded materials

Tutorials of Shibori link1, link2

Licence and credits

Attribution — ShareAlike CC BY-SA This activity has been designed by Anastasia Pistofidou for shemakes.eu. It is based on Fabricademy´s learning content of Biofabricating Materials, FabTextiles´ open publications and Iaac Fab Lab Barcelona´s learning experiences methodology.

Related and supporting activities/modules
It can be combined with the other topics within the Sustainability Package of Fabricademy, such as Biochromes and Textile Scaffold. See clubs, Bacterial Cellulose Leather, Kombucha


Last update: December 23, 2022