用于线虫药物筛选的微流控梯度芯片和自动化平台
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摘要
作为一种在微米尺度操作流体的技术,微流控芯片具有反应体系小、通量高、自动化且操作灵活等优势,被越来越多地应用于细胞和微米尺度生物的研究中。秀丽隐杆线虫作为一种重要的模式生物被广泛地用于神经生物学、衰老及发育和药物筛选等研究中。提出了一种用于研究线虫和环境毒素相互作用的微流控自动化平台,该平台集成了基于微振荡原理的快速梯度形成的微流控芯片、自动化控制系统及基于OpenCV的线虫长度及摆动频率估计的自动化图像分析软件。通过染料和荧光实验验证了基于振荡原理的快速梯度形成芯片,该芯片可以在7 min内形成线性浓度梯度,并通过该芯片和平台验证了线性浓度梯度的双氧水对秀丽隐杆线虫活性的影响。
As a technology for operating fluids on the micrometer scale,microfluidic chips have the advantages of small reaction system,high throughput,automation and flexible operation,and are increasingly used in the research of cells and microscale organisms.As an important model organism,the C.elegans are widely used in the research of neurobiology,aging,development and drug screening.The microfluidic automation platform for the study of the interaction between C.elegans and environmental toxins was presented.The platform integrated the fast gradient forming microfluidic chip based on the micro-oscillation principle,automatic control system and OpenCV-based software for automatic image analysis of C.elegans length and wobble frequency estimation.The fast gradient forming chip based on the oscillation principle was verified by dye and fluorescence experiments.The chip can form a linear concentration gradient within 7 min.The effect of the linear concentration gradient hydrogen peroxide on the activity of C.elegans was verified by the chip and platform.
As a technology for operating fluids on the micrometer scale,microfluidic chips have the advantages of small reaction system,high throughput,automation and flexible operation,and are increasingly used in the research of cells and microscale organisms.As an important model organism,the C.elegans are widely used in the research of neurobiology,aging,development and drug screening.The microfluidic automation platform for the study of the interaction between C.elegans and environmental toxins was presented.The platform integrated the fast gradient forming microfluidic chip based on the micro-oscillation principle,automatic control system and OpenCV-based software for automatic image analysis of C.elegans length and wobble frequency estimation.The fast gradient forming chip based on the oscillation principle was verified by dye and fluorescence experiments.The chip can form a linear concentration gradient within 7 min.The effect of the linear concentration gradient hydrogen peroxide on the activity of C.elegans was verified by the chip and platform.
引文
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[2] CHRONIS N,ZIMMER M,BARGMANN C I.Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans[J].Nature Methods,2007,4(9):727-731.
[3] KATO S,XU Y,CHO C E,et al.Temporal responses of C.elegans chemosensory neurons are preserved in behavioral dynamics[J].Neuron,2014,81(3):616-628.
[4] ALBRECHT D R,BARGMANN C I.High-content behavioral analysis of Caenorhabditis elegans in precise spatiotemporal chemical environments[J].Nature Methods,2011,8(7):599-605.
[5] LARSCH J,VENTIMIGLIA D,BARGMANN C I,et al.High-throughput imaging of neuronal activity in Caenorhabditis elegans[J].Proceedings of the National Academy of Sciences of the United States of America,2013,110(45):E4266-E4273.
[6] JEON N L,DERTINGER S K W,CHIU D T,et al.Generation of solution and surface gradients using microfluidic systems[J].Langmuir,2000,16(22):8311-8316.
[7] DERTINGER S K W,CHIU D T,JEON N L,et al.Generation of gradients having complex shapes using microfluidic networks[J].Analytical Chemistry,2001,73(6):1240-1246.
[8] JUNG J,NAKAJIMA M,TAJIMA H,et al.A microfluidic device for the continuous culture and analysis of Caenorhabditis elegans in a toxic aqueous environment[J].Journal of Micromechanics and Microengineering,2013,23(8):085008-1-085008-8.
[9] WEN H,YU Y,ZHU G,et al.A droplet microchip with substance exchange capability for the developmental study of C.elegans[J].Lab on a Chip,2015,15(8):1905-1911.
[10] DING X T,NJUS Z,KONG T,et al.Effective drug combination for Caenorhabditis elegans nematodes discovered by output-driven feedback system control technique[J].Science Advances,2017,3(10):eaao1254-1-eaao1254-7.
[11]曹哲.基于液滴微流控SDA芯片的粒子合成研究[D].上海:上海交通大学,2017.
[12] CHENG D T,YU Y,HAN C,et al.A simple microdevice for single cell capture,array,release,and fast staining using oscillatory method[J].Biomicrofluidics,2018,12(3):034105-1-034105-2.
[13] ZIVKOVIC Z,van der HEIJDEN F.Efficient adaptive density estimation per image pixel for the task of background subtraction[J].Pattern Recognition Letters,2006,27(7):773-780.
[14] WANG D Y,XING X J.Assessment of locomotion behavioral defects induced by acute toxicity from heavy metal exposure in nematode Caenorhabditis elegans[J].Journal of Environmental Sciences,2008,20(9):1132-1137.