微流体衍射相位显微成像及其在寄生虫测量中的应用

Microfluidic diffraction phase microscopy and its application in parasites measurement

本文提出了一种将衍射相位显微技术与微流体芯片相结合的方法对水源性寄生虫进行定量测量。结合干涉技术与光学显微镜搭建了衍射相位显微成像系统,实现对寄生虫的高灵敏度实时测量。基于光刻工艺,设计和制作了U型捕获结构双层微流体芯片,实现高通量的单个寄生虫捕获。将与聚二甲基硅氧烷(PDMS)折射率相同的聚蔗糖水溶液通入微腔,消除U型捕获结构边缘衍射在相位成像时产生的显著干扰噪声。利用不同直径的标准聚苯乙烯微球验证了该系统的准确性,最大相位值误差不超过3%。采用上述系统测量了100个贾第鞭毛虫包囊和100个隐孢子虫卵囊,然后从干涉图中重构出两虫的相位图。通过对定量相位图的分析得出两虫的形态学参数与定量的光体积差分布,定量的数据为了解其生理特性提供了依据。微流体衍射相位显微成像系统结构简单,稳定性好,测量精度高,在对单个微生物进行实时监测和无标记定量研究方面具有巨大的潜力。

This paper proposes a method of using diffraction phase microscopy combined with microfluidic chip to quantitatively measure waterborne parasites. A diffraction phase microscopy system is built up by combining interferometry with optical microscope to achieve high sensitivity real-time measurement of parasites. Based on soft lithographic techniques, a double-layered microfluidic chip with U-shaped trapping structures is designed and fabricated for high throughput single parasites trapping. Ficoll solution with the same refractive index as polydimethylsiloxane(PDMS) is introduced into the microfluidic chamber to eliminate significant artifacts in phase imaging originating from diffraction at the edges of trapping structures. The accuracy of the system is verified using standard polystyrene microspheres of different diameters, and the error of maximum phase shift does not exceed 3%. 100 Giardia Lamblia(G. Lamblia) cysts and 100 Cryptosporidium Parvum(C. Parvum) oocysts are measured using this system. The phase maps of the parasites are obtained from the interferograms. The morphological parameters and quantitative optical volume difference distribution of the two kind of waterborne parasites are obtained by analyzing the quantitative phase maps. Quantitative data provides the basis for understanding their physiological characteristics. The microfluidic diffraction phase microscopy system has simple structure, good stability and high measurement accuracy, and has great potential for real-time monitoring and label-free quantitative studies of single microorganism.