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Configurable and Up-Scalable Microfluidic Life Science Platform for Cell Based Assays by Gravity Driven Sequential Perfusion and Diffusion

Dimov, Ivan Krastev (2010) Configurable and Up-Scalable Microfluidic Life Science Platform for Cell Based Assays by Gravity Driven Sequential Perfusion and Diffusion. PhD thesis, Dublin City University.

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Abstract

Microfluidics has the potential to significantly change the way modern biology is performed, but for this potential to be realized several on-chip integration and operation challenges have to be addressed. Critical issues are addressed in this work by first demonstrating an integrated microfluidic tmRNA purification and real time nucleic acid sequence based amplification (NASBA) device. The device is manufactured using soft lithography and a unique silica bead immobilization method for the nucleic acid micro purification column. The integrated device produced a pathogen-specific response in < 3 min from the chip-purified RNA. Further enhancements in the device design and operation that allow the on-chip integration of mammalian cell handling and culturing produced a novel integrated NASBA array. This system demonstrated for the first time that it is possible to combine on a single micro-device cell culture and real time NASBA. In order to expand the cell based assay capabilities of the integrated NASBA array and simplify the device operation novel hydrodynamics and cell sedimentation within trench structures and gravity driven sequential perfusion and diffusion mechanisms were developed. These mechanisms were characterized and implemented within an iCell array device. iCell array can completely integrate cell based assays with bio-analytical read-out. The device is highly scalable and can enable the configurable on-chip integration of procedures such as adherent and non-adherent cell-culture, cellstimulation, cell-lysis, cell-fixing, protein-immunoassays, bright field and fluorescent microscopic monitoring, and real time detection of nucleic acid amplification. The device uses on-board gravity driven flow control which makes it simple and economical to operate with dilute samples (down to 5 cells per reaction), low reagent volumes (50 nL per reaction), highly efficient cell capture (100% capture rates) and single cell protein and gene expression sensitivity. The key results from this work demonstrate a novel technology for versatile, fully integrated microfluidic array platforms. By multiplexing this integrated functionality, the device can be used from routine applications in a biology laboratory to high content screenings.

Item Type:Thesis (PhD)
Date of Award:29 March 2010
Refereed:No
Supervisor(s):Ducrée, Jens and Lee, Luke
Uncontrolled Keywords:microfluidics; lab-on-a-chip; µTAS; lab-in-a-trench; cell handling
Subjects:Biological Sciences > Biotechnology
Physical Sciences > Analytical chemistry
Biological Sciences > Biochemistry
Physical Sciences > Nanotechnology
Biological Sciences > Immunology
Biological Sciences > Cell biology
Biological Sciences > Molecular biology
Physical Sciences > Detectors
Medical Sciences > Cancer
Physical Sciences > Chemistry
Biological Sciences > Biosensors
DCU Faculties and Centres:Research Initiatives and Centres > Biomedical Diagnostics Institute (BDI)
DCU Faculties and Schools > Faculty of Science and Health > School of Physical Sciences
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License
Funders:SFI
ID Code:15334
Deposited On:05 Apr 2011 11:01 by Prof. Dr. Jens Ducrée. Last Modified 07 Oct 2013 13:14

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