College - Author 1

College of Engineering

Department - Author 1

Biomedical Engineering Department

Degree Name - Author 1

BS in Biomedical Engineering

College - Author 2

College of Engineering

Department - Author 2

Biomedical Engineering Department

Degree - Author 2

BS in Biomedical Engineering

College - Author 3

College of Engineering

Department - Author 3

Biomedical Engineering Department

Degree - Author 3

BS in Biomedical Engineering

Date

3-2022

Primary Advisor

Christopher Heylman, College of Engineering, Biomedical Engineering Department

Abstract/Summary

The following document outlines the entire process for designing and manufacturing a Live Cell Imaging Gas Control subsystem in tandem with the Heated Stage subsystem to make a CO2 and temperature-controlled chamber for use with the Olympus IX73P2F Microscope. It highlights the current literature and patents surrounding live cell imaging and touches on the existing live cell imaging setups currently being used by other entities. Following this, it defines the scope of the project, key customer requirements, and engineering specifications to ensure the design meets all the needs of its users. This document includes project planning steps to establish important deadlines and a budget for project completion. As the project has progressed, additional documentation has been added such as morphologies of possible design functions, conjoint analyses, conceptual evaluations, and conceptual models for development of the final design. To note, these initial designs highlighting a drawer like chamber were soon dismissed as it would not allow imaging within the device due to objective focal length. The design evolved into the final chamber with a removable lid of a polysulfone chamber with aluminum base. The manufacturing process instructions and design history file show the exact steps taken to manufacture the chamber prototype. Similarly, circuitry diagrams and instructions on the Arduino controlled CO2 system are outlined. In depth test plans were outlined, including testing for the materials, chamber itself, CO2 control system, and fully combined system with the Heated Stage subsystem. Material testing showed that polysulfone, glass, and aluminum withstand autoclave sterilization and laboratory cleaning agents. Chamber testing highlighted the compact size and usability of the device with a multitude of culture vessels. CO2 control system testing showed that the control system effectively keeps CO2 concentration between 4.5% and 5.5% during imaging, with the system being able to re-establish CO2 concentration within 2 minutes of the lid removal, and the system establishes required CO2 concentration about 5 minutes after setup. The entire Live Cell Imaging Device was shown to keep cells alive compared to an ambient air flask for a period of 3 hours. Finally, this document outlines the next steps for this device, including finer tuning of the CO2 control system and increasing cellular viability in the chamber for longer periods of time with the addition of humidity control.

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