Micromachined Multi-cell-line Device For Cell/particle Counting and Sorting Utilizing Image Detection System

• Main Authors: Shen-Chie Chang, 張勝傑
• Other Authors: Gwo-Bin Lee
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/mjns72

碩士 === 國立成功大學 === 工程科學系碩博士班 === 92 === 　　This paper reports a new micromachine-based flow cytometer capable of parallel processing of cell/particle counting and sorting using microfluidic technology. Hydrodynamic focusing of multiple sample streams is achieved simultaneously by using a new layout of sheath flow channels coming form one single inlet port. Thus only one syringe pump is required during the operation. The flow rate of each sheath flow could be much more stable and uniform with this approach. The images of the focused multiple samples are recorded downstream utilizing a high-speed digital CCD camera. Digital image processing technique is then used to count the number of the cells/particles and measure the speed of the cells/particles at the same time. Moreover, the size of the cell/particle can be identified using the image detection system. Theoretical models based on a “flow-rate-ratio” method are used to predict the width and the position of the multiple focused streams, which could be incorporated with digital image system. Experimental data are found to be highly consistent with theoretical results. Subsequently, cell/particle sorting could be achieved using dielectrophoretic (DEP) forces generated by built-in micro-electrodes downstream. The proposed multi-cell-line flow cytometer is fabricated in low-cost soda-lime glass using a rapid fabrication process. Experimental data show that the developed device can successfully detect the number of the cells/particles in two parallel sample streams and the velocities of the cells/particles in each cell line as well. Low counting error and high counting reliability are also verified by counting a mixture of polystyrene beads with different sizes. Human red blood cells are also used for cell counting test. Moreover, cell/particle counting has been demonstrated successfully. The throughput and the performance of the cell counting/sorting device could be greatly improved due to the development of the proposed methods.