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Optofluidic biosensors - miniaturized multi-color flow cytometer and fluorescence-activated cell sorter (microFACS)

机译:光电流体生物传感器-微型多色流式细胞仪和荧光激活细胞分选仪(microFACS)

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We report a portable, low-cost, and high-performance microfluidics based fluorescence-activated cell sorter (microFACS) system to isolate E.coli. cells in combination with a modified specific fluorescence labeling method called tyramide signal amplification-fluorescence in situ hybridization (TSA-FISH). One of the primary challenges in studying bacterial communities that elude cell culturing is to isolate of low abundance bacteria cell from heterogeneous microbial samples. The proposed TSA-FISH protocol is flow cytometry compatible and yields about 10-fold enhancement in fluorescence labeling intensity over widely used standard FISH staining methods. Teflon AF coated optofluidic waveguide and space-time coding with a matched filter algorithm enhance its detection sensitivity. The microFACS is also able to enrich TSA-FISH labeled E.coli. cells by a factor of 223 with an integrated piezoelectric actuator and realtime control electronics system. The microFACS in conjunction with the modified TSA-FISH technologies demonstrates a highly effective and low cost solution potentially for the genomic complexity of complex bacterial communities.
机译:我们报告了一种便携式,低成本,高性能的基于微流体的荧光激活细胞分选仪(microFACS)系统来分离大肠杆菌。细胞与改良的特异性荧光标记方法(称为酪酰胺信号放大-荧光原位杂交(TSA-FISH))结合。研究无法进行细胞培养的细菌群落的主要挑战之一是从异质微生物样品中分离低丰度细菌细胞。拟议的TSA-FISH方案与流式细胞仪兼容,并且与广泛使用的标准FISH染色方法相比,荧光标记强度提高了约10倍。特氟隆AF涂层的光流体波导和带有匹配滤波器算法的时空编码增强了其检测灵敏度。 microFACS还能够富集TSA-FISH标记的大肠杆菌。单元具有集成压电执行器和实时控制电子系统的223倍。结合改进的TSA-FISH技术的microFACS展示了一种高效且低成本的解决方案,潜在地解决了复杂细菌群落的基因组复杂性问题。

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