Title page for ETD etd-10292010-133201


Type of Document Dissertation
Author Dharmasiri, Udara R. Dharmasiri Rasika
Author's Email Address udaraud1@gmail.com
URN etd-10292010-133201
Title Highly Efficient Selection, Enumeration, Enrichment, and Molecular Profiling of Low-Abundance Biological Cells
Degree Doctor of Philosophy (Ph.D.)
Department Chemistry
Advisory Committee
Advisor Name Title
Soper, Steven Committee Chair
Garno, Jayne Committee Member
Spivak, David Committee Member
Zhang, Donghui Committee Member
Francis, Joseph Dean's Representative
Keywords
  • surface modifications
  • Bio-pathogens
  • E.coli
  • Bacteria
  • Antibody
  • Aptamers
  • Conductivity
  • Stem Cells
  • Microfluidics
  • Circulating Tumor Cells
Date of Defense 2010-09-29
Availability unrestricted
Abstract
After brief overviews of low-abundance cell selection techniques in chapter 1 and circulating tumor cells in chapter 2, this dissertation initially focuses on the development of aptamer incorporated high-throughput microfluidic techniques to select rare circulation prostate cancer cells (LNCaP) directly from whole blood with subsequent quantification of these rare cells using a non-labeling approach. Then, I extended the technology to environmental samples in an effort around time, sensitivity, and portability of traditional groundwater assessment. As a model bio- pathogen, E. coli O157:H7 was chosen due to its toxicity and its adverse impact on recreational waters. Low-abundance (<100 cells mL-1) E. coli O157:H7 cells were isolated and enriched from environmental water samples using a microfluidic chip that its capture beds were covalently decorated with E.coli O157:H7 specific polyclonal antibodies. The selected cells were enumerated using RT-qPCR technique. Finally, I have integrated HTMSU with electrokinetic enrichment microfluidic unit for performance of single recombinant low-abundance CTC cell-based assay. A series of analytical processes were carried out, including immunoaffinity selection of rare CTCs, quantification of selected cells via conductivity impedance and electrophoretic enrichment of selected cells for PCR/LDR/CE interrogation for detection of low-abundance point mutations in genomic DNA.
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