Using microfluidic impedance cytometry to measure C. elegans worms and identify their developmental stages
Using microfluidic impedance cytometry to measure C. elegans worms and identify their developmental stages
复制标题
使用微流控阻抗细胞术测量线虫并确定其发育阶段
DOI：
10.1016/j.snb.2018.07.169
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发表时间：
2018-12-01
影响因子：
8.4
通讯作者：
Pan, Dejing
中科院分区：
文献类型：
--
作者：
Zhu, Zhen;Chen, Weijie;Pan, Dejing
微流控阻抗细胞仪允许无标记检测细胞。但它还没有被用于检测大型生物，如秀丽隐杆线虫（C。elegans）蠕虫，由于其多变的形态和强大的运动性。在这里，我们报告一个C。elegans微流控阻抗细胞仪（CeMIC），它能够测量蠕虫流过直微通道时的电阻抗，并根据阻抗信号识别它们的发育阶段。通过对阻抗传感结构的优化配置，即微通道和集成微电极，可以消除蠕虫的起伏和位置变化对阻抗测量的影响。在信号处理中，采用核密度估计方法来提取蠕虫长度相关值，这些值可以直接分配给蠕虫的发育阶段。基于阻抗的蠕虫阶段识别的准确率可达90%。此外，CeMIC设备被开发成一个简单的自动化系统，用于基于大小的蠕虫富集。在通过阻抗测量确定其大小后，成功地将来自混合种群的大小蠕虫分离到不同的出口。因此，我们的CeMIC系统提供了一个很有前途的平台，以测量蠕虫的大小，确定发育阶段，并准备大小/阶段均质化群体的C。elegans实验
Microfluidic impedance cytometry allows for label-free detection of cells. But it has not yet been used to detect large organisms, such as Caenorhabditis elegans (C. elegans) worms, due to their variable morphology and strong motility. Here, we report on a C. elegans microfluidic impedance cytometry (CeMIC), which enables the electrical impedance measurement of worms when flowing through a straight microchannel and the identification of their developmental stages based on impedance signals. With the optimized configuration of impedance-sensing structures, namely the microchannel and integrated microelectrodes, the influence of undulation and position variation of worms upon the measured impedance can be eliminated. In signal processing, a kernel density estimation method is employed to extract worm-length-related values that can be directly assigned to the developmental stages of worms. The accuracy for impedance-based identification of worm stages can reach 90%. Additionally, the CeMIC device is developed into a simple and automated system for size-based enrichment of worms. Large and small worms from a mixed population are successfully separated to different outlets after identifying their sizes with impedance measurement. Therefore, our CeMIC system provides a promising platform to measure the worm size, identify developmental stages, and prepare size/stage-homogenized populations for C. elegans experiments.