基于培养池阵列微流控芯片的单线虫并行分离条件考察

Investigation on Conditions for Parallel Isolation of Single Nematode Based on Microfluidic Chip with an Array of Culture Chambers

秀丽隐杆线虫是一种重要的模式生物,已广泛应用于生物医药、农业和植物方面的研究,但线虫体态微小,常规方法难以实现对单条线虫的精准操控和长期追踪。本研究基于微流体控制技术,设计了培养池阵列微流控芯片,提出一种单线虫并行分离方法。通过考察液体培养时的线虫密度、线虫体宽及其分布等影响因素,分析在芯片上进行单线虫并行分离的条件。结果显示,采用无菌液体培养时,线虫密度对线虫体宽及其分布具有明显影响,高密度(约700条/mL)培养组和低密度(约300条/mL)培养组的线虫体宽分别为(35.5±5.2)μm和(40.0±1.8)μm,且低密度下培养的线虫可获得较好的芯片分离效果,所得含线虫与含单线虫培养池的比例分别为83.3%和66.7%。该并行分离方法相对简便、可靠,结合其微流控芯片结构和流体控制方式,有望用于自动化单线虫长期培养和观测研究。

Caenorhabditis elegans(C.elegans)is an important model organism,which is widely used in biomedical,agricultural and botanical research.However,nematodes are too tiny to accurately manipulate and track single nematode for a long period by conventional methods.Based on microfluidic technology,a microfluidic chip with an array of culture chambers was designed and a method for parallel isolation of single nematode was proposed.The conditions of on-chip parallel isolation of single nematode were analyzed by investigating the factors affecting nematode body width and its distribution in liquid culture.The results showed that the nematode density had a significant effect on the nematode body width and its distribution when it was cultured in a sterile liquid medium.The body widths of nematodes cultured in high density(about 700 worms/mL)and in low density(about 300 worms/mL)were 35.5μm±5.2μm and 40.0μm±1.8μm,respectively.The nematodes cultured in low density could be used to obtain better isolation of single nematode on the chip,and the ratios of the culture chambers containing nematodes and containing single nematode were 83.3%and 66.7%,respectively.This study would provide basic data for further optimization of conditions for single nematode isolation,and wouldlay the foundation for the construction of automated microfluidic systems for parallel culture and long-term observation of single nematode.