Identifier
etd-10292010-133201
Degree
Doctor of Philosophy (PhD)
Department
Chemistry
Document Type
Dissertation
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.
Date
2010
Document Availability at the Time of Submission
Release the entire work immediately for access worldwide.
Recommended Citation
Dharmasiri, Udara R. Dharmasiri Rasika, "Highly efficient selection, enumeration, enrichment, and molecular profiling of low-abundance biological cells" (2010). LSU Doctoral Dissertations. 3081.
https://digitalcommons.lsu.edu/gradschool_dissertations/3081
Committee Chair
Soper, Steven
DOI
10.31390/gradschool_dissertations.3081