1. Live Cell Imaging Chamber
Client: Lance Rodenkirch
Advisor: Prof. John Webster
Jon Sass
David Schurter
Dan Jonovic
Matt Kudek
Bryan Fondrie

2. Design Statement
Engineer a cell imaging chamber so that substances can be perfused into the cell culture at a controlled rate and also maintain a specific carbon dioxide concentration.

3. Live Cell Imaging Two Types Light field Dark field 3-D imaging
Develop studies to establish natural behavior
Induced change to study effects of a controlled factor
Light field
Dark field
3-D imaging

4. Live Cell Imaging Purpose of a Petri dish:
Fluid containment to regulate cell viability
Flat transparent surface for imaging
Isolate cells for manipulation/observation

5. Live Cell Imaging Goal: Imitate cell’s natural environment
Nutrient filled serum
5% CO2 concentration
Temperature set at 37° C
Saturated humidity
Accomplished through:
Perfusion
CO2 regulators
Heating elements
Water pan

6. Design Constraints
Must not interfere with normal functions of microscope
Designed around a 35 mm glass bottom dish
Chamber must have internal access
Microscope platform 21cm x 25cm x 10cm
pH limiting factor, constant 5% CO2
Humid environment
Costs < $1000

7. Design #1: CO2 Feedback and Monitoring System
Enclosed Plexiglas chamber
CO2 sensor provides [CO2] information
Circuit evaluates the relayed information
Adjustments made automatically by CO2 uptake controlling valves
Add CO2 if [CO2] < 5%
Small fan keeps air mixture homogeneous

8. Proposed CO2 Monitoring and Feedback System
Fan
CO2 Sensor
CO2 IN
Air Out
Circuit analyzes sensor voltage
[CO2] higher than desired (unlikely)
[CO2] less than desired
Add CO2
Air IN
Add Air

9. Design #1 Analysis PROS CONS Accurate CO2 monitoring
“Set it and forget it”
Independent of Petri-dish
Rather complicated system
Slower feedback time

10. Design #2: Modified Petri Dish
CO2
Petri dish
Pump
Would directly modify a Petri dish to provide a CO2 input and removal system

11. Design #2 Analysis PROS CONS Few Parts/Simple Design Cheaper Solution
Small area to work with
Reuse same top
Low quality of Petri-dish material
Cannot fit CO2 sensor

12. Design #3: CO2 Infused Medium

13. Design #3 Analysis Pros No enclosure Refreshes Medium
Increased experimental diversity
Cons
Cover-slip Out of Focus
Price
No CO2 concentration
monitoring
Waste of medium

14. Design Matrix 8 4 5 7 24 6 2 19 3 DESIGN CO2 Monitor
Practicality (1-10)
Ease of use (1-5)
Durability (1-5)
Cost Efficiency (1-10)
TOTAL
(1-30)
CO2 Monitor 8 4 5 7 24
Modified Petri Dish 6 2 19
CO2 Infused Medium 3

15. Future Work Finalize design selection
Consult client about further specifications/modifications
Transform design to working prototype
Test prototype and modify
Implement product into working environment

16. Questions?