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

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.

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

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

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

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

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

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

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

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

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

Design #3: CO2 Infused Medium

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

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

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

Questions?