Monitoring ROS generation in single cell by optical fiber-based nano-sensor

• Main Author: Ewe, Ko Xing.
• Other Authors: Li Changming
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/16484
• Institution: Nanyang Technological University

This project investigates the nano-sensing ability of optical fiber-based nano-sensor by the detection of Reactive Oxygen Species (ROS) in living smooth muscle cells. The first part of the project discusses the fabrication process of the nano-sensor. Optical fibers were drawn by micropipette puller using “Heat and Pull” technique, producing sharp tapering end measurable in nanometer scale. Silver coating of the tapering end, followed by silanization with APTES and functionalizing with the ROS-dye complete the fabrication of the nano-sensor. The second part discusses the experiments conducted to study the effects of concentration of EDC and NHS (compounds to facilitate activation of ROS-dye) and the length of tapering end in the efficiency of the nano-sensor. Smooth muscle cells and breast cancer cells (MCF-7) are cultured as test samples for ROS detection. Hydrogen peroxide (H2O2) is added to the smooth muscle cells to stimulate ROS production. This facilitates the study of the nano-sensing ability of nano-sensor fabricated by the techniques discussed in the first part. Confocal microscopy is also employed in this study to investigate the effects of H2O2 in ROS production in smooth muscle cells and MCF-7 cells.