Processing and analysis of motion-blur images in light scattering flow cytometry

F. Yuan, Zhiwen Wang, Qiao Liu, Xuantao Su
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Abstract

Light scattering flow cytometry has been demonstrated for label-free particle and cell analysis. The high-throughput imaging of cells or particles in flow cytometry is fundamentally challenging as motion blur may occur for weak-light measurements. Here we perform light scattering measurements on 3.87 μm and 4.19 μm microparticles in diameters with our light scattering flow cytometer. Two-dimensional (2D) light scattering patterns are imaged by a Complementary Metal Oxide Semiconductor (CMOS) sensor. The hydrodynamic focusing effect is studied for better high-throughput measurements. Motion-blur images are obtained at 2.4 mm/s flow rate with 10 ms exposure time, and a deblurring algorithm is adopted for analysis. The experimental 2D light scattering patterns agree well with the Mie theory simulation. Moreover, the number of 3.87 μm and 4.19 μm microparticles in flow can be determined, where the error is less than 5% compared with the theoretical results.
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光散射流式细胞术中运动模糊图像的处理与分析
光散射流式细胞术已被证明用于无标记颗粒和细胞分析。流式细胞术中细胞或颗粒的高通量成像从根本上具有挑战性,因为弱光测量可能会出现运动模糊。本文用光散射流式细胞仪对直径为3.87 μm和4.19 μm的微粒子进行了光散射测量。二维(2D)光散射模式成像由互补金属氧化物半导体(CMOS)传感器。为了更好地进行高通量测量,研究了流体动力聚焦效应。在2.4 mm/s流速下,曝光时间为10 ms,获得运动模糊图像,并采用去模糊算法进行分析。实验得到的二维光散射图与Mie理论模拟结果吻合较好。此外,还可以确定3.87 μm和4.19 μm颗粒在流动中的数量,与理论结果相比误差小于5%。
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