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

Live cell imaging is a method used by scientists to obtain a better understanding of tissue and cell functions through the study and imaging of cellular dynamics, however it is often detrimental to cell cultures due to effects of changing environment on dynamic cellular events. In order to combat the effects of a changing environment, our team has designed, manufactured, and tested an incubation chamber that will control the heat and pH environment that these cells are experiencing throughout the imaging process. This project is sponsored by the BMED microscopy core group to be used with the Olympus IX73P2F inverted fluorescent microscope by undergraduate and graduate research technicians at Cal Poly.

Our team first considered user requirements which were converted into engineering design specifications. These specifications include but are not limited to, the sample temperature variation from the setpoint value, the total size of the chamber, the time to heat up, the clarity, the accuracy of the temperature probe, and the duration of heating. After defining our requirements, our team designed multiple conceptual ideas to meet these requirements, keeping in mind intellectual property assessment and budgetary constraints, and then used a morphology to determine the best concept. When the final design was completed, the chamber was manufactured, according to our manufacturing process instructions. When the manufacturing had been completed the user manual was written, outlining our instructions for use and warnings to address hazards and risks that come with using the chamber.

The chamber was tested to determine if user requirements had been met by comparing data collected to our engineering specifications. Our team tested the sample variation from the setpoint, the time to heat up the chamber, and the accuracy of the temperature probe. It was concluded that the sample matches the setpoint value and our chamber will keep the sample temperature within +/- 0.5 ̊C, which meets our user specifications. After testing our materials and sizing, it was determined that the chamber can withstand 70% isopropyl alcohol cleaning as well as the autoclave process for sterilization. When imaging at 10x magnification, transmittance testing showed no statistical difference in image clarity between the images taken with and without the chamber. When using 4x magnification the image clarity was not visibly different. Cell samples that were left out on the counter compared to those in the chamber over 3 hours were analyzed under a microscope and indicated less cell death in the samples that were in the device, as expected. The live cell imaging incubation chamber successfully sustains an heated environment that will keep cells viable throughout the imaging process, and does not impede image quality.

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