THESIS
2019
xiv, 81 pages : color illustrations ; 30 cm
Abstract
Microfluidic is a technique for precisely controlling and manipulating microscale fluids,
an interdisciplinary field including engineering, physics, chemistry, micromachining and
bioengineering, etc. Microfluidic chip is the main platform and device for implementing
microfluidic technology, its main feature is that the effective structure (microchannels or
microchambers) containing the fluid is on the micrometer scale in at least one dimension.
In this thesis, we primarily studied microfluidics and its related microsystems and applied
this novel technology in the field of molecular biology including nucleic acid extraction
and its subsequent detection and quantification. Foremost, we introduced basic concepts
and advantages of microfluidics, and we also elaborated on current mat...[
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Microfluidic is a technique for precisely controlling and manipulating microscale fluids,
an interdisciplinary field including engineering, physics, chemistry, micromachining and
bioengineering, etc. Microfluidic chip is the main platform and device for implementing
microfluidic technology, its main feature is that the effective structure (microchannels or
microchambers) containing the fluid is on the micrometer scale in at least one dimension.
In this thesis, we primarily studied microfluidics and its related microsystems and applied
this novel technology in the field of molecular biology including nucleic acid extraction
and its subsequent detection and quantification. Foremost, we introduced basic concepts
and advantages of microfluidics, and we also elaborated on current materials and methods
used to fabricate microfluidic chips and the manipulation of fluids within microchip.
Next, we presented background information regarding nucleic acid extraction and purification
since it is the fundamental step of all downstream modern molecular biology detections
such as polymerase chain reaction (PCR). Microchip-based nucleic acid extraction
and PCR were emphasised because they are important application of microfluidics. Based on the abovementioned reports, we developed an automated and miniaturized device for
rapid nucleic acid extraction which eliminates external centrifugation and precipitation
equipments and can perform the whole extraction process within 10 minutes. 293T cells
were extraction through the device and the extracted total RNA was verified by real-time
PCR and post gel electrophoresis. Furthermore, we presented a portable microchip real-time
PCR to enable rapid detection of the extracted nucleic acid. Actual samples were
used to justify the performance of our device and the detection limit is as low as 1 pg/μL
with a reaction time of 25 minutes. Microchips were fabricated using silicon and glass, and
we carefully designed the microchip to achieve simple yet effective sample introduction
and sealing method. Finally, we reported a digital PCR microchip for high dynamic range
nucleic acid detection. The silicon and glass based microchips prevents water evaporation
during the PCR reaction, and we adopted passive power source which is capillary force
to implement sample introduction which does not require external cumbersome power
supply and also avoid errors caused by different user operations as well as improve uniformity
of the experiment results.
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