Date of Award

Winter 3-3-2018

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biomedical Engineering

First Advisor

Teresa Murray

Abstract

Custom equipment is often necessary in the laboratory. However, costs for such equipment can be high and efficient systems with lower cost are an advantage. In this work, we showed that a cellular environment to keep cells healthy and viable for five hours on a microscope could be created using FDM printing (Chapter 3). Design criteria were that it maintained correct temperature, high humidity and proper pH to control the environment as discussed in Chapter 2. Results indicated that cells would maintain viability for up to five hours, but redesigns needed to be made for extending viability past five hours.

An imaging enhancement termed PICS (Planar Imaging of Curved Surfaces) was also introduced (Chapter 5). For this project, design criteria were that the device stabilized the tissue for imaging while allowing fluid flow around the tissue and facilitate rotation of the naturally curved tissue with respect to the microscope objective lens. Three further considerations of the PICS device were: (1) that it should not add undue shear stresses to the sample tissue, (2) that there should be no unintentional translation, (3) that it should attach to a standard microscope temperature controller in such a way that it does not interfere with temperature controls and does not move unintentionally. Results indicate successful movement of tissue with respect to imaging plane to facilitate productive image capture.

Finally, in Chapter 7, a microelectrochemical array was used to explore differences between glioma and normal astrocyte glutamate levels. Results showed that glioma and normal astrocyte glutamate uptake can be tracked in real-time and compared using the probe. The distinction between glioma cells and astrocytes relies on three indicators: (1) a signal more stepped like in appearance, closer to that of a calibration curve, coming to a steady baseline faster and having a shorter FDHM, is a first indication of impaired uptake; (2) a signal showing a lower k constant, indicating a slower overall clearance rate, (3) faster rise times corresponding with a slower maximum elimination rate. These corresponding factors together indicate impaired uptake in glioma cells versus normal astrocyte glutamate uptake.

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