基于液滴微流控的壳寡糖植物免疫诱导剂荧光检测

发布时间：2019-07-05 11:25

【摘要】：为了建立基于液滴微流控芯片平台的植物免疫诱导剂的荧光信号检测技术,该文设计制作了具有液滴生成结构的芯片,制备包裹烟草(BY-2)细胞的液滴,并对包裹了细胞的液滴数目进行统计分析。将包裹细胞的液滴孵育一氧化氮探针后使用质量浓度为50μg/m L壳寡糖处理,在荧光显微镜下观测其产生的荧光,利用荧光酶标仪对比采用96孔板和液滴承载的细胞的荧光变化趋势。结果显示,水相流速为100μL/h,油相流速为300μL/h时,微流控芯片产生的液滴尺寸适合包裹细胞,其中液滴包裹单细胞的比例达22.9%。经壳寡糖处理后的细胞在生成的液滴中产生了明显的荧光,且用96孔板和液滴承载的细胞的荧光变化趋势一致。研究结果为开发基于液滴微流控技术的植物免疫诱导剂的高通量筛选平台提供参考。
[Abstract]:In order to establish the fluorescence signal detection technology of plant immune inducer based on droplet microfluidic chip platform, a chip with droplet formation structure was designed and fabricated, and the droplets wrapped in tobacco (BY-2) cells were prepared, and the number of droplets wrapped in the cells was statistically analyzed. The nitric oxide probe was incubated with the coated cell droplets and treated with 50 渭 g / mL chitosan oligosaccharide. The fluorescence produced by the probe was observed under the fluorescence microscope. The fluorescence change trend of the cells carried by 96 well plate and droplet was compared by fluorescence enzyme labeling instrument. The results showed that when the flow rate of water phase was 100 渭 L / h and that of oil phase was 300 渭 L / h, the droplet size produced by microfluidic chip was suitable for wrapping cells, and the proportion of single cell wrapped by droplets was 22.9%. The cells treated with chitosan oligosaccharide produced obvious fluorescence in the droplets, and the fluorescence trend of the cells loaded with 96 well plates and droplets was the same. The results provide a reference for the development of a high throughput screening platform for plant immune inducers based on droplet microfluidic technology.
【作者单位】：大连工业大学食品学院;中国科学院大连化学物理研究所;大连海事大学环境科学与工程学院;
【基金】：国家自然科学基金(31370391) 中科院STS计划(KFJ-SW-STS-143) 大连市人才项目(2016RQ064) 辽宁省教育厅项目(2016J008)
【分类号】：TQ458;O657.3

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