Impedance Measurements of Single Basophils Stimulated by Pollen Allergen

• Main Authors: Chi-Ho Chen, 陳繼禾
• Other Authors: 陳志敏
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/5q8944

碩士 === 國立中興大學 === 機械工程學系所 === 106 === This study investigates the allergic reaction caused by birch pollen based on impedance response as a label-free and non-invasive analysis. In the present experimental study, light lithography techniques for microelectromechanical systems (MEMS) were employed to produce the bio-chip for capturing and measuring single cells. In this study, a basophilic cell line KU812 were cultured with specific birch pollen (Betula verrucosa, Bet v 1) IgE through passive transfer as the sample cells. Then specific antigen (anti-Bet v 1 IgE) were used to stimulate the sensitized single KU812 cells that were captured in the bio-chip for investigating the degranulation phenomenon. Experimental results show that the impedance magnitude for capture of single KU812 cell increases to nearly twice compared with the one before the capturing. As the single sensitized cell was stimulated with birch pollen allergens at a concentration of 1 μg/ml, the impedance was found to decrease about 23%. Time-varying images of the stimulated single KU812 cell also shows a rapid expansion in size in about 15 min and then followed by a slower growth until 45 min, resulting in a total increase in cell diameter from 13.3 to 16.1 μm. Using a much higher birch pollen concentration of 10 μg/ml for stimulation also demonstrated a decrease in impedance by about 24%, indicating indiscernible change as compared with the lower concentration of 1 μg/ml. The bio-chip developed in the present experimental study allows one to measure and observe the allergic reaction of IgE-mediated single basophils stimulated by the specific allergens and may serve as a platform for allergy testing.