| 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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| Files |
| Filename |
Size |
Approximate Download Time
(Hours:Minutes:Seconds)
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| 28.8 Modem |
56K Modem |
ISDN (64 Kb) |
ISDN (128 Kb) |
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DISSERTATION.pdf |
8.76 Mb |
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