Microfluidics for Cell Manipulation and Analysis

dc.contributor.authorLoufakis, Despina Nelieen
dc.contributor.committeechairLu, Chang-Tienen
dc.contributor.committeememberBaird, Donald G.en
dc.contributor.committeememberDavalos, Rafael V.en
dc.contributor.committeememberDavis, Richey M.en
dc.contributor.departmentChemical Engineeringen
dc.date.accessioned2014-10-22T08:00:51Zen
dc.date.available2014-10-22T08:00:51Zen
dc.date.issued2014-10-21en
dc.description.abstractMicrofluidic devices are ideal for analysis of biological systems. The small dimensions result to controlled handling of the flow profile and the cells in suspension. Implementation of additional forces in the system, such as an electric field, promote further manipulation of the cells. In this dissertation, I show novel, unique microfluidic approaches for manipulation and analysis of mammalian cells by the aid of electrical methods or the architecture of the device. Specifically, for the first time, it is shown, that adoption of electrical methods, using surface electrodes, promotes cell concentration in a microchamber due to isoelectric focusing (IEF). In contrast to conventional IEF techniques for protein separation, a matrix is not required in our system, the presence of which would even block the movement of the bulky cells. Electric field is, also, used to breach the cell membrane and gain access to the cell interior by electroporation (irreversible and reversible). Irreversible electroporation is used in a unique, integrated microfluidic device for cell lysis and reagentless extraction of DNA. The genomic material is subsequently analyzed by on-chip PCR, demonstrating the possible elimination of the purification step. On the other hand, reversible electroporation is used for the delivery of exogenous molecules to cells. For the first time, the effect of shear stress on the electroporation efficiency of both attached and suspended cells is examined. On the second part of my dissertation, I explore the capabilities of the architecture of microfluidic devices for cell analysis. A simple, unique method for compartmentalization of a microchamber in an array of picochambers is presented. The main idea of the device lies on the fabrication of solid supports on the main layer of the device. These features may even hold a dual nature (e.g. for cell trapping, and chamber support), in which case, single cell analysis is possible (such as single cell PCR). On the final chapter of my dissertation, a computational analysis of the flow and concentration profiles of a device with hydrodynamic focusing is conducted. I anticipate, that all these novel techniques will be used on integrated microfluidic systems for cell analysis, towards point-of-care diagnostics.en
dc.description.degreePh. D.en
dc.format.mediumETDen
dc.identifier.othervt_gsexam:3727en
dc.identifier.urihttp://hdl.handle.net/10919/50586en
dc.publisherVirginia Techen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectmicrofluidicsen
dc.subjectmammalian cellsen
dc.subjectcell focusingen
dc.subjectelectrical cell lysisen
dc.subjectcell electroporationen
dc.subjectchamber formationen
dc.subjecthydrodynamic focusingen
dc.titleMicrofluidics for Cell Manipulation and Analysisen
dc.typeDissertationen
thesis.degree.disciplineChemical Engineeringen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.leveldoctoralen
thesis.degree.namePh. D.en
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