Integrated Cell Lysis and Microfluidic Separation of Erythrocytic Hemoglobin from Blood

• Main Authors: Tsung-Sheng Hung, 洪宗聖
• Other Authors: Shou-Wen Tsai
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/63255945209126644574

碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 98 === A microfuidic device design which integrated photolithography, electroporation, ion-exchange chromatography and photopolymerization technologies to achieve quick cell disruption and subsequent cellular components purification was demonstrated in this research. Comb patterned Pt electrodes were carefully aligned with 40 μm gap by photolithography to extend the cell-disrupting area within the microchannel. Erythrocytes were used as a sample for observing the electrolysis. Various combinations of electropotential, frequency and specialized waveform were examined to optimize the cell disruption. Cell disruption could be observed in the range 35-40 volt/cm and 25-1500 kHz with designed waveform applied to comb electrodes in seconds. Furthermore, these selected parameters are not only avoided bubble formation, but also maintained electrodes from lift-off damage. Target hemoglobin in lysed cellular mixtures was further separated by ion-exchange chromatography. Discontinuous NaCl gradient was used to separate the cellular proteins and subsequently verified by multi-wavelength spectrophotometry. Successful separation shows the potential of this platform as the micro-total-analytical-system (μTAS) for future biological study.