Biological cell manipulation and analysis is one of the most investigated applications of microfluidics. In the last decade, researchers have developed means to handle and sort cells, isolate and study single cells, assay whole and lysed cells, and transfect and electroporate in microchannels. Much of this work was motivated by the observation that many external forces and fields scale favorably in the micro-regime; this is especially the case for the electrical field. This dissertation investigates further integration of electrical forces with microfluidic devices, both channel- and droplet-based, in order to generate new, flexible and more efficient tools for studying cell biology.
The first part of the dissertation (Chapter 3) explores a new dielectrophoretic particle separation method in microchannels. Current electrodeless dielectrophoretic (DEP) separation techniques utilize insulating solid obstacles in a direct current (DC) or low-frequency alternating current (AC) field, while this novel method employs an oil droplet acting as an insulating hurdle between two electrodes. Since the size of the droplet can be dynamically changed, the electric field gradient, and hence DEP force, becomes easily controllable and adjustable to various separation parameters. Very effective separation at the low field strength suggests that this method can also be applied to a separation of biological cells that are not sensitive to low electric potential.
The second, larger part of the dissertation (Chapters 4 and 5) is focused on digital microfluidics (DMF), which is used to actuate nanoliter droplets of reagents and cells on a planar array of electrodes. It was demonstrated for the first time that DMF can be used as a method for cell culture and analysis. Several cell-based applications were implemented in DMF format including long-term culture, cell passaging, assaying and transfection. The data presented here suggest advanced performance of DMF techniques relative to standard macro-scale techniques. Cell analysis using DMF was found to be advantageous because of greatly reduced reagent and cell use, increased sensitivity, and the potential for multiplexing. Also, DMF technique for cell passaging demonstrated faster and more straightforward manipulation of cells than the standard techniques. In addition, no adverse effects of actuation by DMF were observed in assays for cell viability, proliferation, and biochemistry.
The new DMF platform for long-term mammalian cell culture represents the first microfluidic implementation of any kind of all of the steps required for mammalian cell culture – cell seeding, growth, detachment, and re-seeding on a fresh surface. In addition, it is the first demonstration of long-term cell culture in nanoliter droplets. Cells handled in this manner exhibited growth characteristics and morphology comparable to those cultured in standard tissue culture vessels. We anticipate that the DMF cell culture and analysis techniques presented here will be useful in myriad applications that would benefit from automated mammalian cell culture.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OTU.1807/26448 |
| Date | 07 March 2011 |
| Creators | Barbulovic-Nad, Irena |
| Contributors | Wheeler, Aaron |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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