Microfluidic electroporation and cell arrays
Abstract
In this thesis, I present two novel microfluidic techniques for cell-based high throughput analysis: flow-through electroporation and micro-bioreactor arrays. This thesis falls into two main parts. Part I (chapter 2 to 7) demonstrates the electroporative technique and part II (chapter 8 to 9) presents the micro-bioreactor array with individually addressable unit. The flow-through electroporation is achieved by narrowing specific section(s) of microchannels. The narrow section(s) is able to amplify the electric field intensity due to its high resistance. Both transient and permanent electroporations are successfully conducted using this technique. The transient electroporation is witnessed by the cell size expansion and delivery of membrane impermeant molecules into CHO-K1 cells. The fast release of pre-loaded fluorogenic dye from CHO-K1 cells indicates the permanent disrupture of cell membrane under extreme electroporation conditions. A proof-of-concept device for chemical cytometry is also demonstrated using the flow-through electroporation. The effects of electric parameters and buffer concentrations to the electroporation are investigated in detail and optimization of the electroporation is also addressed. The flow-through electroporation can as well be applied to pasteurization when resulting field intensity in the narrow section(s) is larger than 1KV/cm. The micro-bioreactor array is built based on multi-layer soft lithography. The array was designed to provide refined sample maneuver and enclosed microenvironment. Microvalves around each cell culture unit are in charge of the traffics of solutions in the device. Controllable cell seeding (with different amounts and types) and independent treatments of cells in different unit are achieved through actuating microvalves. This micro-bioreactor array overcomes some of the major problems encountered in analysis of multiple samples on the microfluidic platforms.
Degree
Ph.D.
Advisors
Lu, Purdue University.
Subject Area
Biomedical research|Chemical engineering
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