PCR amplification of single cell isolation via droplet-based microfluidics

• Main Author: Hong, Fion Li Ying
• Other Authors: Kang Yue Jun
• Format: Final Year Project
• Language: English
• Published: 2014
• Subjects: DRNTU::Engineering
• Online Access: http://hdl.handle.net/10356/60805
• Institution: Nanyang Technological University

Conventional DNA profiling work involved using a large amount of white blood cells obtained from the crime scene blood sample without quantitation. Therefore, introducing the concept of droplet-based microfluidics to perform single cell encapsulation serves as a potential methodology for DNA quantitation in forensic science applications. The project involved fabricating a lab-on-a-chip device for single cell encapsulation through complete soft lithography process. The cells were stained and visualized under the fluorescence microscope to observe and detect single cell encapsulated within individual droplet. The key factors to achieving the objectives of this study were optimization of oil phase composition, flow rates of both oil and water phases, channel dimensions, and cell densities. These techniques investigated above will lay grounds in developing a droplet-based microfluidic lab chip that is capable of performing single cell isolation for DNA quantitation. By developing a single cell analytical instrument for DNA profiling with the integration of cell lysis and PCR, this would establish a promising foothold for future work in the forensic science field.