Phiphattanaphiphop Chalinee, Leksakul Komgrit, Wanta Thananut, Khamlor Trisadee, Phattanakun Rungrueang
Industrial Engineering Department, Faculty of Engineering, Chiang Mai University, Chiang Mai 50200, Thailand.
Animal and Aquatic Science Department, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand.
Micromachines (Basel). 2022 Mar 10;13(3):426. doi: 10.3390/mi13030426.
This study proposes a microfluidic device used for X-/Y-sperm separation based on monoclonal antibody-conjugated magnetic beads, which become positively charged in the flow system. Y-sperms were selectively captured via a monoclonal antibody and transferred onto the microfluidic device and were discarded, so that X-sperms can be isolated and commercially exploited for fertilization demands of female cattle in dairy industry. Therefore, the research team used monoclonal antibody-conjugated magnetic beads to increase the force that causes the Y-sperm to be pulled out of the system, leaving only the X-sperm for further use. The experimental design was divided into the following: Model 1, the microfluid system for sorting positive magnetic beads, which yielded 100% separation; Model 2, the sorting of monoclonal antibody-conjugated magnetic beads in the fluid system, yielding 98.84% microcirculation; Model 3, the sorting of monoclonal antibody-conjugated magnetic beads with sperm in the microfluid system, yielding 80.12% microcirculation. Moreover, the fabrication microfluidic system had thin film electrodes created via UV lithography and MWCNTs electrode structure capable of erecting an electrode wall 1500 µm above the floor with a flow channel width of only 100 µm. The system was tested using a constant flow rate of 2 µL/min and X-/Y-sperm were separated using carbon nanotube electrodes at 2.5 V. The structure created with the use of vertical electrodes and monoclonal antibody-conjugated magnetic beads technique produced a higher effective rejection effect and was able to remove a large number of unwanted sperm from the system with 80.12% efficiency.
本研究提出了一种基于单克隆抗体偶联磁珠的用于X/Y精子分离的微流控装置,该磁珠在流动系统中带正电。Y精子通过单克隆抗体被选择性捕获并转移到微流控装置上然后被丢弃,这样X精子就可以被分离出来并用于满足乳业中母牛的受精需求。因此,研究团队使用单克隆抗体偶联磁珠来增加使Y精子被拉出系统的力,只留下X精子以供进一步使用。实验设计分为以下几种:模型1,用于分选阳性磁珠的微流系统,其分离率为100%;模型2,在流体系统中分选单克隆抗体偶联磁珠,微循环率为98.84%;模型3,在微流系统中对单克隆抗体偶联磁珠与精子进行分选,微循环率为80.12%。此外,制造的微流系统具有通过紫外光刻制作的薄膜电极以及能够在距底部1500 µm处竖起电极壁且流动通道宽度仅为100 µm的多壁碳纳米管电极结构。该系统以2 µL/min的恒定流速进行测试,并使用2.5 V的碳纳米管电极分离X/Y精子。利用垂直电极和单克隆抗体偶联磁珠技术构建的结构产生了更高的有效排斥效果,能够以80.12%的效率从系统中去除大量不需要的精子。