School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, 510006, China.
Biomed Microdevices. 2013 Apr;15(2):211-20. doi: 10.1007/s10544-012-9719-7.
Behavioral Caenorhabditis elegans mutants are sought for the purposes of neurobiological research. Until now, large numbers of worms with neuronal defects have been obtained through mutagenesis techniques. However, the existing screening procedures are not only time-consuming and low-throughput, but also tedious and labor-intensive. Therefore, developing a rapid and convenient method to overcome these difficulties is necessary. The present study demonstrates for the first time a microdevice for the rapid screening of chemotaxis-defective mutants based on their chemotactic response. The microchip is capable of automatic introduction, local immobilization, and controllable generation of concentration gradients during the screening assays. With this device, six C. elegans behavioral assays can be performed using various attractants without requiring anesthetics for local capture, and ten mutants are effectively isolated from 10(4) mutagenized worms in 100 min. The microfluidics-based method is robust enough to sort the chemotaxis-defective worms with 91 % accuracy from a large population of wild type animals during a mutagenesis screen.
正在寻找具有行为缺陷的秀丽隐杆线虫突变体，用于神经生物学研究。到目前为止，已经通过诱变技术获得了大量具有神经元缺陷的线虫。然而，现有的筛选程序不仅耗时、通量低，而且繁琐、劳动强度大。因此，开发一种快速便捷的方法来克服这些困难是必要的。本研究首次展示了一种基于线虫趋化性反应的快速筛选趋化性缺陷突变体的微器件。该微芯片能够在筛选过程中自动引入、局部固定和可控地产生浓度梯度。使用该装置，无需局部捕获麻醉剂，即可使用各种趋化剂进行六种秀丽隐杆线虫行为测定，并且可以在 100 分钟内从 104 个诱变线虫中有效分离出 10 个突变体。基于微流控的方法足够稳健，可以从大量野生型动物中以 91%的准确率对趋化性缺陷的线虫进行分类，从而实现诱变筛选。
