USMA-West_Point
Notebook
Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.
What should this page have?
- Chronological notes of what your team is doing.
- Brief descriptions of daily important events.
- Pictures of your progress.
- Mention who participated in what task.
Inspiration
You can see what others teams have done to organize their notes:
- AM - Angela Marsh
- JG - Dr. Joel Gaston
- KP - Kanak Patel
- MM - Matthew McDonough
- PB - Payton Boylston
- RG - Rayonna Gordon
- WO - Winston Ou
Perfusion System with Bioreactor Design
Programming and Electronics (Bioreactor focus)
Neurons
9 Jun 2017WO RG AM KP MM
- Received HT-22 Immortalized Mouse Hippocampal Cell Line and related materials from West Point
WO RG AM KP MM PB
- Unfroze neurons
- Plated neurons for amplification
WO RG AM KP MM PB
- Change media
JG
- Changed media
WO RG MM PB
- Split cells - Passage 1 from receipt of neurons
WO RG MM PB
- Changed media
- Froze excess cells
JG
- Changed media
WO RG
- Made and changed media
WO RG
- Thawed and plated cells - Passage 2
WO RG
- Split cells - Passage 3
WO RG
- Changed media
WO RG
- Split cells - Passage 4
WO RG
- Changed media
JG
- Cells were killed
WO RG
- Plated Passage 2 HT22 cells that were previously frozen into a 6 well plate
- Coated MEA with polylysine
- Seeded 20,000 HT22 cells onto coated MEA
- Incubated said cells
- Began bioreactor design based on former models to be used in a perfusion system
- Used AutoDesk Inventor to design a general bioreactor
- Incorporated shape and general items such as holes for screws and cavity for neuronal culturing
- Determined location of a multielectrode array (MEA) in bioreactor design, where the neurons would be transfected and tested
- Adjusted design of bioreactor design
- Began a custom design for a MEA
- Continued MEA designing: location of electrodes, leads, contact pad spacing, materials
- MEA
- Altered the electrode arrangement and size for a single well of a 96 well plate
- Predetermined specifications: Gold or TiN wires, embedded in glass
- Determined custom MEA design specs, particularly spacing
- Revised MEA spacing
- Discussed the location of holes for media flow and a bubble trap
- Fleshed out design for bubble trap
- Began design for clamp to keep pins on the multielectrode array (extension of bioreactor) - KP
- Began discussion on removal of olfactant contaminated media while maintaining a perfusion system via syringes
- Continued clamp design-KP
- Requested a quote for custom MEA
- Confirmed the use of a peristaltic pump for media flow
- Finalized MEA design
- Contacted other MEA fabrication companies for quotes
- Continued search of MEA vendors
- Worked on specifics regarding flow into and out of the bioreactor/media reservoirs - decided to use luer locks
- Continued contacting MEA vendors
- Determined best luer lock connectors for tubing
- Finished printing of clamp design
- Explored photomasking vendors
- Determined design of media reservoirs to be conical flasks
- Explored pre-made MEA vendors
- Determined design of media reservoirs to be conical flasks
- Explored pre-made MEA vendors
- Ordered MEAs from Multichannel Systems
- Researched on systems to control the peristaltic pump from the computer
- Find tubing for bioreactor and pump
- Designed media reservoirs in Inventor, obtaining a cubic appearance, as opposed to the original conical tube appearance
- Ordered tubing
- Redesigned clamp system for pins using a platform and screws system
- Revised bioreactor design to have internal channels in one side of the bioreactor
- Inserted a conical steeple for the bubble trap
- Made minor modifications to bioreactor design
- Inserted threading into media reservoirs
- Made measurements of ordered o-rings to be fitted
- Printed out models to help test fittings for the O-ring
- Printed array to narrow down sizing for O-Ring
- Modified media reservoir due to space constraints
- Printed model MEAs to test pin setup and general fittings
- Modified media reservoir for better fluid flow
- Printed first part of bioreactor
- Adjusted dimensions and reprinted first part of bioreactor
- Adjusted dimensions and printed a third version of the first part of bioreactor
- Printed array to determine necessary dimensions to fit luer locks
- Applied dimension changes to bioreactor design and reprinted the first part of bioreactor
- Determined screw size and lengths needed to apply appropriate pressure on pins
- Printed out top and bottom of bioreactor and clamp
- Perforated MEA Bioreactor modified
- Verified operation of the pump
- Tested calibration of pump
- Printed bioreactor top in Dental SG and media reservoir in clear
- Tested perfusion system with bioreactor and MEA to test for leakage and overall flow
- Printed bioreactor bottom in Dental SG and waste reservoir in clear
- Revised bioreactor design for better hole sizing and placement
- Tested perfusion system with bioreactor, media reservoir, and MEA to test for leakage and overall flow
- Redesigned reservoir for more efficient drainage
- Printed bioreactor top in Dental SG
- Tested perfusion system with all components to test for leakage and overall flow
- Printed both reservoirs in Dental SG
- Tested perfusion system with Dental SG bioreactor top/bottom and MEA to test for leakage and overall flow
- Cured bioreactor pieces in UV
- Performed dry run with cured pieces
- Autoclaved epoxy-containing piece
- Autoclaved entire perfusion/bioreactor system
- Introduced to labview - use to take voltage readings from electrodes
- Determined to set peak threshold to +/- 2 SD of base activity
- Built time-averaging method to determine frequency and amplitude of peaks
- Generated preliminary statistics for data
- Built code to write to a file
- Inserted method to adjust thresholds before data collection
- Adjusted labview program to read in data from electrodes using Intan codes (as opposed to theoretical inputs)
- Finished labview program - can select filename/type
- Implemented impedance program from Intan to determine the presence of cells on electrodes
- Soldered pins to electrode adapter boards
- Tested program that ran impedance tests and voltage readings
- Soldered more electrode adapters
Perfusion System with Bioreactor Design
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Programming and Electronics (Bioreactor focus)
23 May 2017WO
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