Please note that this is a draft schedule and will inevitably change to some extent.

1 | 8/28 | Starter Cultures

In Class

  • Welcome
  • Lecture: What is Biofabrication?
  • Website Overview
  • Syllabus
  • Video: Suzanne Lee
  • Introduction to the Bio Lab
    • EHS
    • Communication
  • Lab Work: K.hansenii Starter Culture for growing Bacterial Cellulose
    • Work in teams of two students
    • Make HS Media
      • Each team will make 1L of media
    • Inoculate 10 mL cultures
      • 1 culture per student
      • one negative control per team

Homework



2 | 9/04 | Culture Growth

In Class

  • Make HS Media
    • Work in pairs, each team will make 1L of media
  • Discussion
  • Presentation
    • Lecture – Biofabrication with Bacterial Cellulose
  • Lab
    • examine 10mL K. Hansenii cultures
      • check to see if there is a pellicle
      • note the thickness of the pellicle
      • snap a photo of your culture, focused on the pellicle at the top
    • scale up K. hansenii cultures
      • transfer the culture, including the pellicle, to a sterile 25mL conical tube
      • vortex the tube to distribute the cells into the media
        • set the vortexer switch to off (O) and the dial to 2000 RPM.
        • snap two 25mL tubes (with lids tightly closed) into the vortexer
        • set a timer for 20 seconds
        • switch the vortexer on (II) for 20 seconds
      • using a 200 uL micropipette, pull 100uL of culture from the 25mL conical tube and dispense it into a 1.5mL microfuge tube (for serial dilution exercise)
      • Add to ~ 200mL of HS media to a sterile 500mL Erlenmeyer flasks
      • Add all of the remaining culture and the pellicle from the 25mL conical tube to the Erlenmeyer flask
      • Secure the sponge closure on the Erlenmeyer flask
      • Place the flask in an incubator set to 28C
    • discuss next scale up to 1-5L
    • serial dilution lab exercise to determine cell culture density
    • look at cells and cellulose under the microscope

Homework

  • Lab
    • Serial Dilution
      • check on plates
        if you cannot make these times, ask a classmate to photograph your plates
        once you have readable plates, no further check-in are required
        additional dates will be scheduled if needed
        • Times TBD
      • once plates are readable
    • K. hansenii Cultures
      • begin planning your bacterial cellulose project
      • make HS media
        • determine volume needed, based on the desired size pellicle and needed vessel
        • make media and autoclave it
        • consolidate your media as soon as possible to free up bottles
  • Reading
  • Prepare for next week’s lab session:


3 | 9/11 | Scaling Up

In Class

Homework


4 | 9/18 | CRISPR Part I – Classroom

In Class

Homework

  • check on K. hansenii cultures
  • review CRISPR reading from last week
    • complete exercises, if not already done
  • preview CRISPR Lab
  • review pipetting videos (accurate pipetting with very small volumes will be critical for CRISPR)
  • begin planning 2D Bioprinting Project

5 | 9/25 |CRISPR Part II – Lab

In Class

Homework

  • check on CRISPR plates
    • Times TBD
  • begin planning and preparing for 2D Bioprinting Project
    • gather tools
    • create stencils
    • prepare files

6 | 10/02 | Bioprinting Experimentation

In Class

  • review prepared materials
  • Demos
    • making agar
      • standard and black
      • with and without antibiotics
    • pouring agar plates
    • manual application of cells
    • ultrasonic vapor deposition
    • Grasshopper for laser-cut stencils
    • Grasshopper bioprinter
    • Bioprinter demo
  • Experimentation

Homework

  • check on experimental plates
    • observe and document growth as frequently as possible
    • remove stencils as needed
    • move plates to refrigerator as needed
  • continue planning and preparing for 2D Bioprinting Project
    • gather tools
    • create stencils
    • prepare files
    • make agar plates

7 | 10/09 | Bioprinting Work 1

In Class

  • Lab work day
  • Demo
    • preserving plates
    • others as needed

Homework

  • check on plates
    • observe and document growth as frequently as possible
    • remove stencils as needed
    • move plates to refrigerator as needed
    • preserve as needed
  • continue work on 2D Bioprinting Project

8 | 10/16 | Bioprinting Work 2

In Class

  • Lab work day
  • Demo
    • preserving plates
    • others as needed

Homework

  • check on plates
    • observe and document growth as frequently as possible
    • remove stencils as needed
    • move plates to refrigerator as needed
    • preserve as needed
  • continue work on 2D Bioprinting Project

9 | 10/23 | Work Day Bioprinting Project – Cellulose Harvest

In Class

Homework

  • Finish bioprinting project
  • document project
  • complete washing protocol
    • This process involves daily steps. Share the labor with classmates as needed to make this manageable
  • complete cellulose treatment samples

10 | 10/30 | Cellulose Conditioning and Work Day

In Class

  • Critique
  • Demo on cellulose treatment methods
  • Distribute clean BC samples for testing
  • class ends 12:30

Homework

  • complete bacterial cellulose project

11 | 11/06 | Critique Bacterial Cellulose Project

In Class

  • 9:00-9:30 Install
  • 9:30 Critique
  • presentation: mycelium in art, design, and manufacturing
  • Introduce Mycelium Project

Homework

  • upload your bacterial cellulose project documentation using the provided template
  • prepare a mold for mycelium casting -or- bring a pattern ready for vacuum forming

12 | 11/13 | Mycelium Inoculation

In Class

  • vacuum forming molds
  • prepare substrates for mycelium project
  • autoclave mycelium substrates

Homework

  • autoclave mycelium substrates
  • inoculate substrates with mushroom cultures
  • reading:



13 | 11/20 | Thanksgiving

14 | 11/27 | MycoCasting

In Class

  • fill molds with inoculated substrate

Homework


15 | 12/04 | Growing Networks


16 | 12/11 | Critique Mycelium Project