Song Pengfei, Dong Xianke, Liu Xinyu
Department of Mechanical Engineering, McGill University , Montreal, Quebec H3A 0C3, Canada.
Biomicrofluidics. 2016 Feb 26;10(1):011912. doi: 10.1063/1.4941984. eCollection 2016 Jan.
The nematode worm Caenorhabditis elegans has been widely used as a model organism in biological studies because of its short and prolific life cycle, relatively simple body structure, significant genetic overlap with human, and facile/inexpensive cultivation. Microinjection, as an established and versatile tool for delivering liquid substances into cellular/organismal objects, plays an important role in C. elegans research. However, the conventional manual procedure of C. elegans microinjection is labor-intensive and time-consuming and thus hinders large-scale C. elegans studies involving microinjection of a large number of C. elegans on a daily basis. In this paper, we report a novel microfluidic device that enables, for the first time, fully automated, high-speed microinjection of C. elegans. The device is automatically regulated by on-chip pneumatic valves and allows rapid loading, immobilization, injection, and downstream sorting of single C. elegans. For demonstration, we performed microinjection experiments on 200 C. elegans worms and demonstrated an average injection speed of 6.6 worm/min (average worm handling time: 9.45 s/worm) and a success rate of 77.5% (post-sorting success rate: 100%), both much higher than the performance of manual operation (speed: 1 worm/4 min and success rate: 30%). We conducted typical viability tests on the injected C. elegans and confirmed that the automated injection system does not impose significant adverse effect on the physiological condition of the injected C. elegans. We believe that the developed microfluidic device holds great potential to become a useful tool for facilitating high-throughput, large-scale worm biology research.
线虫秀丽隐杆线虫因其生命周期短且繁殖力强、身体结构相对简单、与人类有显著的基因重叠以及易于培养/成本低廉,已被广泛用作生物学研究中的模式生物。显微注射作为一种将液体物质输送到细胞/生物体对象中的成熟且通用的工具,在秀丽隐杆线虫研究中发挥着重要作用。然而,秀丽隐杆线虫显微注射的传统手动操作过程劳动强度大且耗时,因此阻碍了涉及每天对大量秀丽隐杆线虫进行显微注射的大规模秀丽隐杆线虫研究。在本文中,我们报告了一种新型微流控装置,该装置首次实现了对秀丽隐杆线虫的全自动、高速显微注射。该装置由片上气动阀自动调节,可对单个秀丽隐杆线虫进行快速加载、固定、注射和下游分选。为进行演示,我们对200条秀丽隐杆线虫进行了显微注射实验,结果表明平均注射速度为6.6条/分钟(平均线虫处理时间:9.45秒/条),成功率为77.5%(分选后成功率:100%),均远高于手动操作的性能(速度:1条/4分钟,成功率:30%)。我们对注射后的秀丽隐杆线虫进行了典型的活力测试,并确认自动注射系统不会对注射的秀丽隐杆线虫的生理状况造成显著不利影响。我们相信,所开发的微流控装置具有巨大潜力,有望成为促进高通量、大规模线虫生物学研究的有用工具。