Gu Xin, Huang Wei, Yang Limei, et al. Microfluidic diffraction phase microscopy and its application in parasites measurement[J]. Opto-Electronic Engineering, 2019, 46(12): 190046. doi: 10.12086/oee.2019.190046
Citation: Gu Xin, Huang Wei, Yang Limei, et al. Microfluidic diffraction phase microscopy and its application in parasites measurement[J]. Opto-Electronic Engineering, 2019, 46(12): 190046. doi: 10.12086/oee.2019.190046

Microfluidic diffraction phase microscopy and its application in parasites measurement

    Fund Project: Supported by National Natural Science Foundation of China (61505240), the Applied Basic Research Programs of Suzhou City (SYG201414), and the Youth Innovation Promotion Association of CSA (2015258)
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  • 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.
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  • Overview: Quantitative phase imaging (QPI) is developed based on light microscopy as an advanced modality for label-free biomedical optical imaging. Diffraction phase microscopy (DPM) is demonstrated as a novel QPI technique, which combines many of the best attributes of current QPI methods. In the quantitative phase imaging process, the suspended sample is usually placed between a coverslip and a glass slide for observation imaging. The sample is easy to aggregate and form clusters and have weak Brownian motion in this case, which can introduce unexpected noise interference in the imaging area.

    In this paper, we propose a method of using DPM combined with microfluidic chip to quantitatively measure Giardia Lamblia (G. Lamblia) cysts and Cryptosporidium Parvum (C. Parvum) oocysts. The DPM system is placed at the output port of conventional light microscope. The DPM interferometer is created using a diffraction grating in conjunction with a 4f lens system. A three-dimensional structure of a microfluidic chip is fabricated using polydimethylsiloxane (PDMS). The chip consists of parallel arrays of U-shaped trapping structures, which contained between 4 and 5 traps over its width. Each row of traps is placed at the gap from the previous row to allow the sample to be fully trapped. A double-layered structure is designed and fabricated to increase trap efficiency and reduce pressure in the chip. 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%. The microfluidic phase imaging system can be accurately used for quantitative phase imaging. 100 G. Lamblia cysts and 100 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.

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    沈阳化工大学材料科学与工程学院 沈阳 110142

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