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
- Eac
h team will make 1L of media
- Eac
- Inoculate 10 mL cultures
- 1 culture per student
one negative control per team
Homework
- Website
- if your Google profile picture is not a recognizable image of yourself, please change it
- log in to the class website, using the “continue with google” option
- confirm that your photo shows up on the the class People page
- if needed, you can make changes to your “display name” on your profile page
- Lab
finish making media (if needed)- check on cultures for cellulose growth -> pellicle formation -> thickness of 2mm+
check on Negative Control
- Reading
- Prepare for next week’s lab session:
- Review the HS Media protocol
- Review units of measurement for the bio lab
- Watch pipetting tutorial videos (~7 minute)
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
- examine 10mL K. Hansenii cultures
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
- count the colonies on the plates
- photograph the plates
- log the data in the spreadsheet
- check on plates
- 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
- Serial Dilution
- Reading
- Prepare for next week’s lab session:
- Review units of measurement for the bio lab
- Review pipetting tutorial videos as needed
3 | 9/11 | Scaling Up
In Class
- Lab
- examine serial dilution exercise results
- review plates
- spreadsheet data
- discuss results
- examine 200mL K. hansenii cultures
- scale up K. hansenii cultures to 1-5L
- look at K. hansenii and cellulose under the microscope
- examine serial dilution exercise results
- Lecture
- Discussion
- Donna Haraway’s Anthropocene, Capitalocene, Plantationocene, Chthulucene: Making Kin
- Synthetic Biology: A Primer, Ch 1
Homework
- Lab
- Scale up K. hansenii cultures to 1-5L
- Read (printouts are in the cubbies just outside of the lab)
- CRISPR lab guide pgs 1-10
- Do worksheet exercises in Part I
- Synthetic Aesthetics Ch1 “Synthetic Biology: What It Is and Why It Matters” by Alistair Elfick and Drew Endy
4 | 9/18 | CRISPR Part I – Classroom
In Class
- Lecture:
- molecular biology for genetic engineering
- Genetic Engineering with CRISPR
- Review CRISPR text and worksheet
- Prepare for CRISPR lab
Discuss Synthetic Aesthetics Ch1 “Synthetic Biology: What It Is and Why It Matters” by Alistair Elfick and Drew Endy- Introduce 2D Bioprinting Project
Lecture: Artists using synthetic biology- check on K. hansenii cultures
Homework
- check on K. hansenii cultures
- review CRISPR reading from last week
- complete exercises, if not already done
- preview CRISPR Lab
- CRISPR lab guide pgs 10-21
- Slide lecture (slides 35-57)
- 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
- Review CRISPR homework
- CRISPR Lab preview
- CRISPR Lab
- check on K. hansenii cultures
- 2D Bioprinting
- Presentation: example work
- Techniques
- direct drawing with various tools
- stamps
- stencils
- hand cut
- laser cut
- image to stencil Grasshopper template
- bioprinter
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
- making agar
- 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
Critique- work day
- Harvest Bacterial Cellulose and begin washing protocol
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
- fill molds with inoculated substrate (if not done in class)
- document process
- reading: A Symbiotic View of Life: We Have Never Been Individual – paper by Scott Gilbert, et. al.
- related recommended readings:
- I Contain Multitudes – book by Ed Yong – Chapter 1 Living Islands
- Ways of Being – book by James Bridle – Chapter 1 Thinking Otherwise, Chapter 2 Wood Wide Web
- Staying with the Trouble – book by Donna Haraway
- related recommended readings: