用于公牛精子细胞单细胞检测和计数的微流控传感器

Chalinee Phiphattanaphiphop, R. Phatthanakun, K. Leksakul
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摘要

介绍了一种具有检测和计数功能的公牛精子检测微流控传感器芯片。该芯片采用紫外光和软光刻工艺制备,以制备电极和微通道,用于细胞对准和输送。此外,该芯片还集成了一对金微电极作为基于lcr阻抗电路的细胞计数器。这种特殊设计的电路的主要目的是减少微电极的大接口阻抗,使其易于使用,以提高计数过程中的精度和效率。然而,精液中精子以外的其他细胞在通过对电极时也会引起阻抗的变化,从而干扰精子计数。这项工作表明,电阻抗的变化与通过电极的细胞的大小有关,从而可以区分精子和其他细胞,即悬浮在洗涤介质中的珠子。工作设计为测量三个样品的平均阻抗;在2 MHz, 2.5V的信号频率下研究培养基,珠粒和精子。平均输出阻抗分别为143.14 kΩ、151.26 k$\Omega$和152.67 k$\Omega$。然后,对这些数据进行归一化处理,比较珠精和牛精信号的差异。所有分析值均显示数据的一致性,p值小于0.05。这项工作提供了重要的信息,因为当精子和小珠流过检测电极区域时,该设备可以利用阻抗作为可区分的细胞计数器来开发。
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A Microfluidic Sensor for Single Cell Detection and Counting Bull Sperms Cell
A microfluidic sensor chip for bull sperm detection with ability of detection and counting is presented. The chip was fabricated by UV and soft lithography process in order to create electrodes and microchannel for cell alignment and delivery. Moreover, the chip was integrated with a pair of gold microelectrodes acted as a cell counter based on LCR-impedance circuit. Main objective of this specially designed circuit is reducing the large interface impedance of the microelectrodes, make them easy to use, for higher accuracy and efficiency in counting process. However other cells than those spermatozoa in semen also cause change of impedance when passing the pair of electrodes and interfere with the spermatozoa count. This work demonstrated that the change in electrical impedance is related to the size of cells passing through the electrodes, allowing distinguishable between spermatozoa and other cells, i.e. beads suspended in washing media. Work has been designed to measure average impedance of three samples; media, beads and spermatozoa investigated at signal frequency of 2 MHz, 2.5V. The averaged output impedances were 143.14 kΩ, 151.26 k$\Omega$, and 152.67 k$\Omega$, respectively. Then, these data were normalized as aspect to compare the differences between signal of beads and bull sperms. All analyzed values show consistencies of data with p-value lower than 0.05. This work provided important information as the device can be developed utilizing the impedance as a distinguishable cell-counter when spermatozoa and beads flow pass the area of detecting electrodes.
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