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首页> 外文会议>Photons Plus Ultrasound: Imaging and Sensing 2019 >Measuring the Nucleus-to-Cytoplasmic Ratio in PC-3 Cells Using Photoacoustic Flow Cytometry and Imaging Flow Cytometry
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Measuring the Nucleus-to-Cytoplasmic Ratio in PC-3 Cells Using Photoacoustic Flow Cytometry and Imaging Flow Cytometry

机译：使用光声流式细胞仪和成像流式细胞仪测量PC-3细胞中的核质比

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摘要

A cell’s nucleus-to-cytoplasm (N:C) ratio is a histological metric used to stage malignant disease. Current N:C assessmentmethods, such as optical microscopy, are time-consuming, subjective, and low-throughput. Here, we compare the N:Cratios of prostate cancer (PC-3) cells measured by a novel microfluidic PhotoAcoustic Flow Cytometer (PAFC) to thoseobtained using an Imaging Flow Cytometer (IFC). PC-3 cells were stained with DRAQ-5 nuclear dye and divided intopopulations measured using the PAFC and IFC. The PAFC consisted of a microfluidic device integrated with a singleelementultrasound transducer (375 MHz central frequency) and a sub-nanosecond pulsed laser (532 nm). Individual cellswere 3D flow-focused through the overlapping focal region of the ultrasound and laser pulses. PAFC estimation of the celland nucleus diameters were determined through power spectra fitting of backscattered US waves and emitted PA waves toestablished theoretical models. An ImageStreamX® IFC was used to acquire brightfield and fluorescent images ofindividual cells, which were masked, gated, and used to assess the cell (brightfield) and nucleus (fluorescence) diameterto validate the PAFC measurements. The average cell and nucleus diameters determined using the PAFC (n = 388) were18.8 ± 3.3 μm and 14.3 ± 2.9 μm, respectively. The corresponding values from the IFC (n = 4651) were 18.3 ± 2.2 μm and12.2 ± 1.9 μm. The N:C ratio (calculated as the ratio of the nucleus diameter to cell diameter) was 0.77 ± 0.10 using thePAFC and 0.67 ± 0.07 using the IFC. Our novel PAFC device has the potential to be used for circulation tumor cell detectionusing the N:C ratios of cells.
机译：细胞核与细胞质（N：C）的比率是用于分期恶性疾病的组织学指标。当前的N：C评估\方法\（例如光学显微镜）是耗时，主观且低通量的。在这里，我们比较了通过新型微流光声流式细胞仪（PAFC）测得的前列腺癌（PC-3）细胞的N：C \ r \ n \ n \ n \ n与使用成像流式细胞仪（IFC）获得的N \ C \ r \ n \ r \ n \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ r \ n \ r \ n \ r \ n的人。用DRAQ-5核染料将PC-3细胞染色，并分成使用PAFC和IFC测量的种群。 PAFC由集成了单元素\超声换能器（中心频率375 MHz）和亚纳秒脉冲激光（532 nm）的微流控设备组成。单个细胞通过超声波和激光脉冲的重叠焦点区域进行3D流动聚焦。通过将反向散射的US波和发射的PA波的功率谱拟合到已建立的理论模型来确定细胞\ r \ n和原子核直径的PAFC估计。 IFCImageStreamX®IFC用于获取单个细胞的明场和荧光图像，对它们进行掩蔽，门控，并用于评估细胞（明场）和细胞核（荧光）直径，以验证PAFC测量。使用PAFC（n = 388）确定的平均细胞直径和细胞核直径分别为\ r \ n18.8±3.3μm和14.3±2.9μm。来自IFC的相应值（n = 4651）为18.3±2.2μm和\ r \ n12.2±1.9μm。使用PAFC时，N：C比（以核直径与细胞直径之比计算）为0.77±0.10，而使用IFC时为0.67±0.07。我们的新型PAFC装置具有利用细胞的N：C比率检测循环肿瘤细胞的潜力。

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  • 来源
    《Photons Plus Ultrasound: Imaging and Sensing 2019》|2019年|108782H.1-108782H.7|共7页
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  • 作者

    Joseph A. Sebastian; Vaskar Gnyawali; Lianne Ho Yan So; Michael J. Moore; Scott S. H. Tsai; Michael C. Kolios;

  • 作者单位

    Department of Physics, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada,Keenan Research Centre, Toronto, Canada;

    Department of Mechanical and Industrial Engineering, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada;

    Keenan Research Centre, Toronto, Canada;

    Department of Mechanical and Industrial Engineering, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada;

    Keenan Research Centre, Toronto, Canada;

    Department of Physics, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada,Keenan Research Centre, Toronto, Canada;

    Department of Mechanical and Industrial Engineering, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada;

    Keenan Research Centre, Toronto, Canada;

    Department of Physics, Ryerson University, Toronto, Canada,Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, Canada,Keenan Research Centre, Toronto, Canada mkolios@ryerson.ca;

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